WO2011096782A2 - Liquid flow apparatus, fixed quantity liquid supply apparatus, and target substance extracting apparatus and target substance extracting method using the two apparatuses - Google Patents

Abstract

The present invention relates to a liquid flow apparatus for conducting tests including biochemical reactions, to a fixed quantity liquid supply apparatus for automatically filling the liquid flow apparatus with liquid substances, to a target substance extracting apparatus for extracting target substances such as nucleic acids, protein or the like from a variety of biological samples, using the liquid flow apparatus and the fixed quantity liquid supply apparatus, and to a target substance extraction method for extracting target substances from a variety of biological samples, using magnetic particles which selectively adsorb target substances. In one embodiment of the liquid flow apparatus according to the present invention, a liquid flow plate comprises: a plurality of storage chambers in which liquid substances are stored; a reaction chamber in which two or more liquid substances react with each other; a plurality of delivery channels interposed between the storage chambers and the reaction chamber so as to deliver the liquid substances stored in the storage chambers to the reaction chamber; injection channels for injecting liquid substances into the plurality of storage chambers; and a discharge channel connected to the reaction chamber, wherein pump means connected to the discharge channel operate and simultaneously the plurality of injection channels are selectively opened/closed, to thereby supply the liquid substances stored in the storage chambers to the reaction chamber, or discharge the liquid substances in the reaction chamber to the outside.

Description

Liquid flow device and liquid metering device, target material extraction device and target material extraction method using the same

The present invention provides a liquid flow device for performing a test including a biochemical reaction, a liquid metering device for automatically filling a liquid material into a liquid flow device, and various biological devices using the liquid flow device and the liquid metering device. A target material extraction apparatus for extracting a target material such as nucleic acid or protein from a sample, and a target material extraction method for extracting the target material from a variety of biological samples using magnetic particles that selectively adsorb the target material.

Recently, as the cause of the disease is analyzed at the genetic level based on the results of research on the human genome, the demand for manipulation and biochemical analysis of biological samples is increasing for the purpose of healing or preventing human diseases.

In addition, in addition to the diagnosis of diseases, technologies for extracting biological materials such as nucleic acids or proteins from biological samples or samples containing cells are required in various fields such as new drug development, examination of viral or bacterial infections, and forensic medicine.

Precipitation, liquid extraction, electrophoresis, chromatography and the like are used to separate biological substances such as nucleic acids and proteins from biological samples. Recently, solid state extraction has been developed to simplify this operation.

The solid phase extraction method is a method using solid particles prepared by attaching a solid particle having a selectivity or a ligand having a high selectivity to a solid phase. After dissolving the biological sample in the reagent solution, the target material is attached to the solid phase, the solid particles attached to the target material are separated from the reagent solution, and the remaining liquid remaining in the solid phase is removed to remove other impurities. Principle is to separate the desired target material from

Representatively US Pat. Nos. 3,970,518 and 3,985,649 disclose methods for separating target materials using fine magnetic particles having large surface areas. In this method, the biochemical material is attached to the magnetic particles while the magnetic particles are dispersed in the reagent solution, a magnetic field is applied to agglomerate the magnetic particles to which the target material is attached, and the remaining solution is removed to separate the target material.

This method is automated and various extraction equipments have been developed. In order to automate this material extraction process, specific manipulation methods such as attaching the target substance, removing and washing the target substance, and separating the target substance should be performed. For example, a mixing operation for uniformly dispersing magnetic particles in a reagent solution containing a biochemical and a flocculation operation for agglomerating magnetic particles dispersed in a reagent solution are basically required.

On the other hand, the operation method of uniformly mixing the magnetic particles into the solution, a method of forming a vortex by shaking the container containing the solution vigorously, or stir the solution with a stick, and the vortex by repeatedly discharging and sucking the solution And a method of forming the same. As an operation of agglomerating magnetic particles dispersed in a solution, a method of basically applying a magnetic field is known. In addition, the position where the magnetic particles are aggregated is changed according to the position of applying the magnetic field. Since the magnetic particle aggregation position is important for efficiently removing the solution, various techniques have been developed regarding the aggregation position of such magnetic particles.

For example, a magnet is placed at the bottom of the container to agglomerate magnetic particles to the bottom of the container, a magnet is placed at the side of the container to agglomerate magnetic particles on the wall of the container, and a donut-shaped magnet is moved. A method of agglomerating magnetic particles in a circle on the inner wall of the tube has been developed. All of these methods aggregate the magnetic particles in a tubular reaction vessel.

In addition, US Pat. No. 6,187,270 discloses a method of aggregating magnetic particles by separating permanent particles from a solution by accessing a permanent magnet to a disposable tip. In this method, magnetic particles are attached to the inside of the tip to separate the magnetic particles out of the reaction vessel. However, this method has a limitation in processing a large number of samples quickly because the tip to which magnetic particles are attached must be moved to several reaction vessels.

In addition, US Patent No. 6,040,192 discloses a method for separating and dispersing magnetic particles while moving between the magnetic rod and the seal that can move up and down between the reaction vessel containing the solution. However, this method also has a disadvantage in that a large amount of disposable seals are consumed and a transfer means for transferring the magnet rods and seals should be provided.

For example, Korean Patent No. 10-0483684 discloses a container partitioned with a plurality of chambers for separating magnetic particles using magnetic rods and seals. This container is arranged in a row of a plurality of chambers with the top open. Therefore, in order to separate and disperse the magnetic particles while moving between the various chambers, a conveying means for moving the encapsulation and the magnet rod up and down while moving left and right must be provided.

Meanwhile, Korean Patent No. 10-0720044 discloses a multi well plate kit. In a multi well plate kit, a plurality of chambers are arranged up, down, left and right. Therefore, in order to separate and disperse the magnetic particles while moving between the various chambers, an extraction apparatus using magnetic rods and seals in a two-dimensional array is required. This has the problem that the device is complicated, large in size and expensive.

In addition, the prior art has to replace the pipette or the tip of the pipette or the container containing the solution and the seal to which the magnetic particles are attached every time the reagent solution is changed, so that not only these consumables are consumed in large quantities, but also the time for changing them This was a lot of troublesome and cumbersome procedure. Recently, a device for automatically inserting a tip or a seal has been developed. However, such a device has a problem of not only complicated structure, taking up a lot of space, but also an increase in equipment price.

The present invention does not consume a large amount of consumables by using a pipette or a tube-shaped container in which a tip or a pipette is inserted at the tip of the pipette, and a seal in which magnetic particles are attached. It relates to a target material extraction device that does not require complicated devices.

The present invention relates to a liquid flow device capable of performing all tests involving biochemical reactions using one plate or block type vessel.

Recently, various kinds of lab-on-a-chips have been developed for analyzing biological samples. Lab-on-a-Chip uses micromaterials such as plastic, glass, and silicon to create microchannels, allowing small amounts of liquid samples to be transferred to perform biological tests. The lab-on-a-chip is basically composed of a controller for controlling the liquid flow between the chamber and the various functions. For example, a storage chamber for storing a sample or a reagent solution, a reaction chamber for reacting a sample and a solution, a moving channel connecting the storage chamber and the reaction chamber to move the fluid, and installed in the moving channel to interrupt the movement of the fluid. It consists of a controller.

In particular, the conventional wrap-on-a-chip allows the liquid channel stored in a plurality of chambers to be selectively supplied by installing a valve in the moving channel. However, the valve installed in the moving channel not only complicates the structure of the lab-on-a-chip but has a problem of limiting the structure of the target material extraction device.

That is, in the conventional wrap-on-a-chip, it is difficult to stack the wrap-on-a-chip up and down or left and right since the valve must be operated through an operation mechanism installed outside the chip. Therefore, the target material extracting apparatus using the conventional wrap-on-a-chip has to arrange a plurality of wrap-on-a-chips horizontally, which increases the size of the apparatus and complicates the structure of the operation mechanism for operating the valves installed in the plurality of the lab-on-a-chips. There is a problem that the size of the device is increased and the price is increased.

In addition, the conventional wrap-on-a-chip has a problem that it takes a lot of trouble and time-consuming in the pretreatment process, because the liquid material to be filled in advance using a pipette in the storage chamber of the chip.

The present invention is to solve the problems according to the prior art, the main object of the present invention, a liquid flow device capable of performing all the tests including the biochemical reaction in one plate or block and liquid to the plate or block By providing a liquid quantitative supply device capable of supplying quantitative substances, it provides a target material extraction device and a target material extraction method that simplify the structure of the target material extraction device and facilitate the target material extraction process including the pretreatment process. It is.

In another aspect, the present invention provides a liquid flow apparatus capable of performing all tests including biochemical reactions in one plate or block and a liquid quantitative supply device capable of quantitatively supplying a liquid substance to the plate or block. The present invention provides a target material extraction device and a target material extraction method capable of extracting a target material from a biochemical sample without using various consumables such as a tip, a seal, and a tube-shaped container attached to the tip of the pipette.

In addition, the present invention stores the liquid material in which the magnetic particles are dispersed in the storage chamber of the liquid flow plate or block, and supply the liquid material in which the magnetic particles are dispersed to the reaction chamber and mixed with other liquid materials to the target material to the magnetic particles The present invention provides a liquid flow apparatus capable of combining and fixing magnetic particles to an inner wall of a reaction chamber to separate a target substance contained in a biological sample, and an apparatus and method for extracting a target substance using the same.

The present invention also provides a liquid flow apparatus capable of storing a plurality of liquid materials, mixing and reacting two or more liquid materials, agglomerating and fixing magnetic materials, and discharging and recovering remaining liquid materials or eluates. will be.

The present invention provides a liquid flow block and a target material extraction apparatus and method using the same, which can automatically process a large amount of samples at once by stacking a plurality of liquid flow plates.

In addition, the present invention is provided with injection channel opening and closing means for selectively opening and closing a plurality of injection channels provided in the liquid flow apparatus, and pump means for forming a vacuum in the reaction chamber or mixing chamber provided in the liquid flow apparatus Not only do not use various consumables such as tip, seal, tube-shaped container, etc. attached to the tip of pipette or pipette, but also omit the pipette or seal transfer device to simplify the structure and the target material extraction device using the same It is to provide an extraction method.

As a means for achieving the above object of the present invention, the liquid flow apparatus according to the present invention largely consists of a liquid flow plate and a liquid flow block.

The liquid flow plate,

A plurality of storage chambers storing liquid materials;

A reaction chamber for reacting two or more liquid materials;

A plurality of feed transfer channels formed between the storage chamber and the reaction chamber for supplying the liquid material stored in the storage chamber to the reaction chamber;

An injection channel formed to inject a liquid material into the plurality of storage chambers;

And a discharge channel formed to be connected to the reaction chamber.

By operating the pump means connected to the discharge channel and selectively opening and closing the plurality of injection channels to supply the liquid material stored in the storage chamber to the reaction chamber or to discharge the liquid material in the reaction chamber to the outside .

The liquid flow plate,

A mixing chamber formed spaced apart from the reaction chamber;

A mixed transfer channel configured to suck the liquid material in the reaction chamber into the mixing chamber or to discharge the liquid material in the mixing chamber into the reaction chamber;

And an air suction channel formed at an upper end of the mixing chamber so as to suck air in the mixing chamber or inject air into the mixing chamber.

By operating the pump means connected to the air suction channel and selectively opening and closing the plurality of injection channels to suck the liquid material stored in the storage chamber into the mixing chamber or to discharge the liquid material of the mixing chamber to the reaction chamber It is characterized by mixing the substances.

The liquid flow plate,

An eluate chamber formed spaced apart from the reaction chamber;

An eluate transfer channel formed to transfer the eluate in the reaction chamber to the eluate chamber;

And an eluate recovery channel formed in the eluate chamber to suck the eluate.

By operating the pump means connected to the eluate recovery channel and selectively opening and closing the plurality of injection channels it characterized in that the eluate stored in the storage chamber to suck or discharge to the eluate chamber.

In the liquid flow plate,

The lower portion of the reaction chamber is characterized in that the installation portion for further allowing the magnetic application means or the heater to be closer.

The liquid flow plate,

A second reaction chamber formed spaced apart from the reaction chamber and connected to the storage chamber through the feed transfer channel;

A second eluate transfer channel formed to transfer the eluate in the reaction chamber to the second reaction chamber;

And a liquid material suction channel formed to be connected to the second reaction chamber.

Operating the pump means connected to the eluate recovery channel and selectively opening and closing the plurality of injection channels to suck the eluate from the reaction chamber and supply the liquid substance stored in the storage chamber to the second reaction chamber for reaction. It is done.

In the second reaction chamber,

A second installation portion for allowing access of the heater or the magnetic application means;

Observation window for observing the reaction inside the second reaction chamber is characterized in that it is further provided.

A plurality of storage chambers for storing a liquid material, an injection channel formed to inject liquid material into the storage chamber, a reaction chamber spaced apart from the storage chamber, and a liquid material stored in the plurality of storage chambers A plurality of feed transfer channels connected to supply the reaction chamber to the reaction chamber, a mixing chamber formed spaced at one side of the reaction chamber, and connected to the reaction chamber through a mixing transfer channel, and connected to a lower end of the mixing chamber to mix In the liquid flow block consisting of a plurality of unit liquid flow plate comprising a discharge channel for discharging the liquid material in the chamber and an air suction channel connected to the upper end of the mixing chamber for discharging the air in the mixing chamber,

The injection channel is formed through the unit liquid flow plate;

The plurality of unit liquid flow plates may be stacked in such a manner that injection channels of neighboring unit liquid flow plates are arranged in the same line.

In the liquid flow block,

The lower portion of the plurality of reaction chamber is characterized in that the installation portion to further enable the magnetic application means or the heater to be close.

The liquid flow block,

A second reaction chamber formed spaced apart from the reaction chamber and connected to the storage chamber through the feed transfer channel;

A second eluate transfer channel formed to transfer the eluate in the reaction chamber to the second reaction chamber;

And a liquid material suction channel formed to be connected to the second reaction chamber.

The liquid flow block,

An eluate chamber formed spaced apart from the reaction chamber;

An eluate transfer channel formed to transfer the eluate in the reaction chamber to the eluate chamber;

And an eluate recovery channel formed in the eluate chamber to suck the eluate.

In the target substance extraction apparatus using the liquid flow plate,

The target material extraction device is mounted to the liquid flow plate,

Injection channel opening and closing means for selectively opening and closing a plurality of injection channels formed in the liquid flow plate;

Pump means connected to the discharge channel, the air suction channel, and the eluate recovery channel formed in the liquid flow plate to suck the liquid substance or the eluate in the reaction chamber, the mixing chamber or the eluate chamber;

Magnetic application means for fixing the magnetic particles to the inner wall of the reaction chamber;

It characterized in that it comprises a control means for controlling the injection channel opening and closing means and pump means and the magnetic application means.

In the target material extraction apparatus using the liquid flow block,

The target material extraction device is mounted to the liquid flow block,

A main body having a mounting portion for mounting the liquid flow block;

Liquid material and air supply means for injecting a liquid material through a plurality of injection channels formed on the side of the liquid flow block in close contact with the liquid flow block mounted on the mounting portion;

Liquid material and air suction means for sucking liquid material and air through a plurality of injection channels formed on the other side of the liquid flow block;

Liquid material conveying and discharging means for discharging the liquid material through the plurality of discharge channels and the air suction channels formed on the other side of the liquid flow block, or mixing the liquid material by sucking or pressurizing air;

Eluent transfer and discharge means for transporting or discharging the eluate through a plurality of eluent recovery channels formed on another side of the liquid flow block.

The target material extraction device using the liquid flow block,

Magnetic application means for agglomerating and fixing magnetic materials on the inner walls of the plurality of reaction chambers;

It characterized in that it further comprises a control means for controlling the magnetic application means, liquid material and air supply means, liquid material and air suction means, liquid material transport and discharge means and eluent transport and discharge means.

The liquid material and air supply means,

A plurality of main supply pipes tightly coupled to a plurality of injection channels formed in the liquid flow block;

A plurality of liquid material tanks for storing the liquid material;

A liquid material supply pipe connecting the main supply pipe and the liquid material tank;

A liquid substance control valve installed in the plurality of liquid substance supply pipes;

A plurality of air supply pipes connected to the plurality of main supply pipes and the other end of which communicates with the outside air;

Installed in the air supply pipe is characterized in that it comprises a plurality of air supply control valve to regulate the flow of air.

The liquid material and air suction means,

A plurality of main suction pipes tightly coupled to a plurality of injection channels formed in the liquid flow block;

A plurality of main pumps installed in the main suction pipe to form a vacuum in the injection channel;

A liquid material drain pipe connected to the main pump to discharge liquid material;

It is characterized in that it comprises a plurality of main control valve is installed in the plurality of main suction pipe to open and close the main suction pipe.

The liquid material transfer and discharge means,

A plurality of liquid material discharge pipes connected to a plurality of discharge channels provided on another side of the liquid flow block;

A main liquid material discharge pipe connected to the plurality of liquid material discharge pipes;

A discharge pump installed at the main liquid material discharge pipe;

A liquid substance discharge control valve installed in the main liquid substance discharge pipe;

A liquid material drain pipe connected to the discharge pump;

A plurality of air discharge pipes connected to the air suction and discharge channels of the liquid flow block;

A main air discharge pipe connected to the plurality of air discharge pipes;

A mixing pump installed at the main air discharge pipe;

An air exhaust pipe connected to the mixing pump;

It characterized in that it comprises an air discharge control valve installed in the main air discharge pipe.

The eluate transfer and recovery means,

A plurality of eluate discharge pipes connected to a plurality of air suction channels provided on another side of the liquid flow block;

A main eluate discharge pipe connected to the plurality of eluate discharge pipes;

An eluent pump connected to the main eluent discharge pipe;

It is characterized in that it comprises an eluent discharge control valve installed in the main eluent discharge pipe to control the discharge of the eluent.

In the target material extraction method for extracting a nucleic acid material from a biological sample using a target material extraction device equipped with a liquid flow plate,

The target material extraction method,

The liquid flow plate filled with the liquid material in the plurality of storage chambers is seated on the mounting unit provided in the target material extraction device, and the valve body of the injection channel opening and closing means provided in the target material extraction device is brought into close contact with a corresponding injection channel. A liquid flow plate mounting step of connecting means to the discharge channel, the air suction channel and the eluate recovery channel of the liquid flow plate;

A liquid material supplying step of controlling the injection channel opening and closing means and the pump means to selectively supply the liquid material stored in the storage chamber of the liquid flow plate to the reaction chamber;

Controlling the injection channel opening and closing means and the pump means to mix the liquid substance supplied to the reaction chamber or to disperse solid particles such as magnetic particles;

A magnetic particle fixing step of agglomerating the dispersed magnetic particles inside the reaction chamber by using the magnetic applying means and attaching the magnetic particles to the inner wall of the reaction chamber;

Controlling the injection channel opening and closing means and the pump means to discharge the remaining liquid in the reaction chamber to the outside;

And controlling the injection channel opening and closing means and the pump means to transfer the eluate in the reaction chamber to the eluent chamber or to recover the eluent to an external storage container through the eluate recovery channel.

In the target material extraction method for extracting a nucleic acid material from a biological sample by using the target material extraction apparatus equipped with the liquid flow block,

The target material extraction method,

The liquid flow that seats the liquid flow block on the mounting portion provided in the main body and in close contact with the liquid material and air supply means, the liquid material and air suction means, the liquid material transfer and discharge means and the eluent transfer and discharge means provided around the mounting portion Block mounting step;

A liquid material filling step of injecting a liquid material into each of the storage chambers through the plurality of injection channels provided in the liquid flow block by using the liquid material and air suction means and the liquid material and air supply means;

A liquid material supplying step of selectively supplying the liquid material stored in the storage chamber of the liquid flow block to the reaction chamber by using the liquid material transport and discharge means and the liquid material and air supply means;

By mixing the liquid material supplied to the reaction chamber by the liquid material transfer and discharge means and the liquid material and air supply means to the mixing chamber or by discharging the liquid material of the mixing chamber into the reaction chamber to mix two or more liquid materials A liquid material mixing step of dispersing solid particles such as magnetic particles;

A magnetic particle fixing step of aggregating the dispersed magnetic particles inside the reaction chamber by using the magnetic application means and fixing the magnetic particles to the inner wall of the reaction chamber;

A liquid material discharge step of discharging the remaining solution inside the reaction chamber and the mixing chamber to the outside by using the liquid material transfer and discharge means and the liquid material and air supply means;

Eluent transfer and recovery step of transferring the eluate in the reaction chamber to the eluent chamber by using the eluent transfer and discharge means and the liquid material and air supply means or recovers to an external storage container through the eluent discharge channel It is characterized by.

The liquid material filling step,

By operating a plurality of main pumps provided in the liquid material and the air suction means to form a vacuum in the main suction pipe, and close the plurality of air control valves provided in the liquid material and the air supply means, and connect with the liquid material storage tank. All the liquid material control valves are opened, and the liquid material of the liquid material storage tank is transferred through the injection channel by a vacuum formed inside the injection channel of the liquid flow block, and the liquid material to be transferred is injected into the injection channel. A liquid material injection step of sequentially introducing a plurality of storage chambers connected to the storage chambers and filling liquid materials into respective storage chambers and injection channels;

By operating a plurality of main pumps provided in the liquid material and air suction means to form a vacuum in the main suction pipe, and at the same time, the plurality of liquid material control valves provided in the liquid material and air supply means are all closed and connected to the outside air. The air control valves of the plurality of air supply pipes are opened to allow external air to be sucked into the main suction pipe through the injection channel, until the vacuum formed in the injection channel is parallel to the surface tension and gravity inside the storage chamber. The liquid material stored in the storage chamber is discharged to the outside through the injection channel by the air injection step of making the level of the liquid material in each storage chamber constant.

Liquid quantitative flow block according to another embodiment of the present invention,

A plurality of injection channels penetrating from one side to the other side of the block;

A plurality of storage chambers installed at predetermined intervals along the injection channel and having a neck portion connected to the injection channel at an upper end thereof;

It is connected to the lower end of the plurality of storage chambers and the end is configured to include a plurality of discharge channels formed to be connected to the outside air,

The liquid material stored in the storage chamber is discharged to the discharge channel by sucking the liquid material through the pump means connected to the injection channel or by sucking or discharging air.

In the liquid quantitative supply device for quantitatively supplying a liquid substance using a liquid quantitative flow block,

The liquid metering device,

Liquid material and air supply means installed on one side of the liquid metering flow block and connected to one side of the injection channel;

And a liquid material and air suction means installed on the other side of the liquid metering flow block and connected to the other side of the injection channel.

The liquid material and air supply means may include a plurality of main supply pipes tightly coupled to a plurality of injection channels formed in the liquid metering flow block, a plurality of liquid material tanks storing liquid materials, and the main supply pipe and liquid material tanks A liquid material supply pipe for supplying a liquid material by connecting a liquid material control valve installed in the plurality of liquid material supply pipes, a plurality of air supply pipes connected to the plurality of main supply pipes and communicating with an outside air, and the air A plurality of air supply control valves installed in the supply pipe to regulate the flow of air;

The liquid material and air suction means, a plurality of main suction pipes are in close contact with the plurality of injection channels formed on the other side of the liquid metering flow block, and are installed to be connected to the main suction pipe to form a vacuum in the injection channel A plurality of main pumps, a liquid material drain pipe connected to the main pump to discharge liquid substances, a plurality of main control valves installed at the plurality of main suction pipes to open and close a main suction pipe, and connected to the main pump It characterized in that it comprises an air discharge pipe for discharging air, and an air discharge control valve installed in the air discharge pipe.

In the liquid quantitative supply method using the liquid quantitative supply device,

The liquid quantitative supply method,

Filling a liquid material into a plurality of injection channels formed in the liquid metering flow block and a plurality of storage chambers connected to the injection channels;

An air suction step of sucking air in the injection channel so as to equalize the level of the liquid material injected into the plurality of storage chambers;

And a liquid material discharge step of discharging the liquid material stored in the storage chamber to the outside.

The liquid material filling step,

By operating the main suction pump connected to the injection channel of the liquid metering flow block to form a low pressure inside the main suction passage and the injection channel, it is installed in the air supply pipe connected to the injection channel of the liquid metering flow block The liquid material control valve installed in the liquid material supply pipe connected to the injection channel is opened while the air control valve is closed, and the liquid material of the liquid material tank flows into the injection channel through the main supply pipe, The injected liquid material is characterized in that it is sequentially filled in the storage chamber connected to the injection channel.

The air suction step,

By operating the main suction pump connected to the injection channel of the liquid metering flow block to form a low pressure in the main suction passage and the injection channel, the air control valve in a state in which the liquid material control valve is closed By opening the air, it is characterized in that the air of the outside air is injected into the injection channel through the main supply pipe to be discharged to the outside through the main suction pipe and the main drain pipe.

In the liquid material discharge step, by operating the main suction pump connected to the injection channel of the liquid metering flow block to form a high pressure in the main suction passage and the injection channel, the liquid material control valve and air By closing the control valve, the high pressure is formed in the storage chamber is characterized in that the liquid material stored in the storage chamber is discharged to the container or multi-well plate kit through the discharge channel.

According to the present invention, all tests involving biochemical reactions can be performed in one liquid flow plate or liquid flow block, thereby automating the process of extracting biological materials such as nucleic acids or proteins from various biological samples such as cells of animals and plants. have.

The present invention also selectively opens and closes injection channels formed in the plurality of storage chambers, and simultaneously forms a vacuum pressure in the reaction chamber or the mixing chamber to selectively supply, mix, and discharge liquid materials, thereby improving the structure of the flow apparatus. In addition to the simplicity, it can be used by stacking a number of plates, there is an effect that can process a large amount of samples at once.

In addition, the present invention can quickly process a large amount of bio information by using a liquid flow block in which a plurality of unit liquid flow plates are stacked vertically, eliminating the need for a plurality of tubular containers and seals or transfer devices for them. In addition to reducing the cost of use, it is possible to reduce the size of the compact.

In addition, the present invention can automate the preparation process by automatically metering various types of liquid material into the storage chamber, and because the magnetic particles are fixed to the inner side wall of the reaction chamber of the liquid flow device, It is possible to reduce the consumption of the consumables because there is no need for sealing, and there is no need for a conveying means for moving the pipette having the tip, the sealing or the magnetic particles fixed up, down, front, rear, left and right, thereby simplifying the structure of the apparatus.

1 is a perspective view showing an example of a liquid flow plate according to the present invention,

2 is a plan view of the liquid flow plate shown in FIG.

3 is a cross-sectional view of the liquid flow plate shown in FIG.

4 is a plan view showing another embodiment of a liquid flow plate according to the present invention;

5 is a cross-sectional view showing another embodiment of a liquid flow plate according to the present invention;

6 is a plan view showing another embodiment of a liquid flow plate according to the present invention;

7 is a perspective view showing an example of an apparatus for extracting a target substance using a liquid flow plate according to the present invention;

8 is a perspective view showing another embodiment of the target material extraction apparatus using a liquid flow plate according to the present invention;

9 is a flowchart showing an example of a method for extracting a target substance according to the present invention;

10 to 16 is an explanatory diagram for showing a target material extraction method according to the present invention,

17 is a perspective view showing a unit liquid flow plate constituting a liquid flow block according to the present invention;

18 is a perspective view showing a liquid flow block according to the present invention,

19 is an explanatory diagram showing an internal configuration of a liquid flow block shown in FIG. 18;

20 is a schematic perspective view showing the structure of a target material extraction apparatus according to the present invention;

21 is an explanatory diagram showing a structure of a target material extracting apparatus according to the present invention;

22 is a plan view for explaining the structure of the target material extraction apparatus according to the present invention;

23 is a flow chart showing a target material extraction method using a liquid flow block of the present invention,

24 to 30 are explanatory views for showing a target material extraction method according to the present invention,

31 is a schematic diagram showing the configuration of a liquid quantitative supply device according to the present invention;

32 is a flowchart showing a liquid quantitative supply method according to the present invention;

33 to 35 are explanatory views showing a liquid quantitative supply method according to the present invention.

Hereinafter, with reference to the accompanying drawings will be described in detail a preferred embodiment of the liquid flow apparatus according to the present invention, the target material extraction apparatus using the same and a method thereof.

First, referring to FIGS. 1 and 4, one embodiment of a liquid flow device according to the present invention is a flat plate-shaped liquid flow plate 10.

The liquid flow plate 10 is formed by bonding two or three thin plates to each other, and there are a plurality of chambers for storing or reacting liquid materials and a plurality of channels for injecting, discharging, or transporting liquid materials. It is provided.

Preferably, the liquid flow plate 10 may be formed by bonding two plates formed with grooves or through portions to each other, or by bonding thin plates to upper and lower surfaces of the plate on which the grooves or through portions are formed.

The liquid flow device of the present invention is similar to the conventional wrap-on-a-chip in that all tests involving biochemical reactions can be performed on one plate, with the difference that no valve is provided in the transfer channel. Therefore, the liquid flow apparatus of the present invention has a feature that can be used by stacking a plurality of liquid flow plate (10).

First, as shown in FIGS. 1 to 3, one embodiment of the liquid flow plate 10 includes a plurality of storage chambers 22, a feed transfer channel 15 and a reaction chamber 25. The storage chamber 22 is for storing a sample, a reagent solution, a solution in which magnetic particles are dispersed, and the like, and a plurality of storage chambers 22 are formed at the upper end of the plate. In addition, each storage chamber 22 is formed with an injection channel 13 for injecting a liquid material connected to the outside, respectively. Preferably, the injection channel 13 may be formed as a through hole penetrating the upper plate 10.

One or more reaction chambers 25 are formed below the storage chamber 22. The reaction chamber 25 is connected to a plurality of storage chambers 22 through a plurality of transport channels, more specifically, a feed transport channel 15.

In the liquid flow plate 10 of the present invention, the feed feed channel 15 has a diameter much smaller than that of the storage chamber 22. For example, the diameter of the channel is 1mm to 5mm. Therefore, the liquid material stored in the storage chamber 22 cannot be transferred to the reaction chamber 25 through the feed transfer channel 15 by itself without external force. In the drawing, a plurality of storage chambers 22 are formed in the same size, but the size of each storage chamber 22 may be formed in a different size depending on the amount of material to be stored.

Then, as shown in Figure 4, the liquid flow plate 10 of the present invention is further provided with a mixing chamber (27). The mixing chamber 27 is installed to be connected to the reaction chamber 25 to function to mix the solution inside the reaction chamber 25.

To this end, the mixing chamber 27 is connected to the reaction chamber 25 through the mixing transfer channel 16. A discharge channel 41 is formed at the lower end of the mixing chamber 27 to be connected to the outside.

In addition, another embodiment of the liquid flow plate 10 may further include an eluent chamber 28. The eluate chamber 28 is connected to the reaction chamber 25 through the eluate transfer channel 17. In addition, an eluate recovery channel 42 is further formed in the eluate chamber 28. Therefore, the target substance (eluate) in the reaction chamber 25 is extracted through the eluate chamber 28.

In addition, an air suction channel 43 is further formed at an upper end of the mixing chamber 27. The air suction channel 43 is connected to the outside air to suck the liquid substance in the reaction chamber 25 and send it to the mixing chamber 27 or conversely pressurize the liquid substance in the mixing chamber 27 and discharge it to the reaction chamber 25. It serves to mix the liquid material.

Preferably, the ends of the eluate recovery channel 42, the discharge channel 41 and the air suction channel 43 is located on the side of the plate, the receiving groove is formed so that the syringe or the pumping means is in close contact. In addition, an injection hole 32 having a rubber stopper 31 for injecting a substance such as a reagent or a sample by using a pipette or the like may be formed at an upper end of some storage chambers 22-1.

5, another embodiment of the liquid flow plate 10 includes upper and lower plates 10 and 11 and an intermediate plate 12 therebetween. As shown, the intermediate plate 12 is formed with a plurality of through portions forming the plurality of storage chambers 22 and the reaction chamber 25. In addition, grooves forming channels are formed on inner surfaces of the upper plate 10 and the lower plate 11. In addition, grooves corresponding to the grooves formed in the upper and lower plates 10 and 11 are formed on the upper and lower surfaces of the intermediate plate 12, respectively, to form supply and transfer channels 15a and 15b. The upper and lower plates 10 and 11 are formed with through holes for forming the injection channel 13 and the discharge channel 41, respectively.

And the intermediate plate 12 is formed with a connecting channel (15c) for connecting the feed transfer channels (15a, 15b) formed on the upper and lower surfaces of the intermediate plate 12, respectively, the liquid material in the connecting channel (15c) A check valve 15d is provided to prevent this backflow. The check valve 15d is formed in the connecting channel 15c penetrating the intermediate plate 12 and blocks the flow of the liquid material flowing through the valve channel 19 and the valve channel formed with the inclined surface to one side. It consists of a check ball 21 to be made. The check ball is installed in a through hole in which a slanted surface is formed to block a liquid material from flowing backward.

Subsequently, referring to FIG. 6, a second reaction chamber 25a is further installed in another embodiment of the liquid flow plate 10 of the present invention. The second reaction chamber 25a is connected to the reaction chamber 25 through the second eluate transfer channel 17a. In addition, a liquid material suction channel 45 is formed in the second reaction chamber 25a to be connected to the outside. Therefore, the eluate of the reaction chamber 25 may be introduced into the second reaction chamber 25a by the pumping action of the pump means installed in the liquid material suction channel 45.

In addition, the second reaction chamber 25a is connected to one or more storage chambers 22 through the feed transfer channel 15. Therefore, the liquid material stored in the storage chamber 22 may be supplied to the second reaction chamber 25a according to the action of the pump means installed in the liquid material suction channel 45.

In addition, an observation window 29 may be further formed on the second reaction chamber 25a to analyze a material in which the reaction occurs in the second reaction chamber 25a, and the second reaction chamber 25a may be formed. In the lower portion of the second reaction chamber (25a) is formed a second mounting portion (18a) for installing a vibrator, a heater, a laser or the like for promoting the reaction in the interior. Reference numeral 29 is an optical analyzer.

Meanwhile, the size of the storage chamber 22 of the liquid flow plate 10 according to the present invention is determined according to the type of liquid material. Some of the plurality of storage chambers 22 are used as sample storage chambers for storing sample solutions. Preferably, an injection hole 32 for injecting a sample solution using a pipette is formed at an upper end of the sample storage chamber. And the injection hole 32 is provided with a rubber stopper 31 to prevent the inflow of air and foreign matter. In addition, although not shown, a recovery port 34 for recovering the eluate may be further formed in the eluate chamber 28. Therefore, the eluate can be recovered by using a pipette or a syringe.

And the feed transfer channel 15 formed in the liquid flow plate 10 is of a size so small that the liquid material stored in the storage chamber 22 does not flow into the feed transfer channel 15 without the help of external force. For example, the diameter of the feed channel 15 is 1mm ~ 5mm does not flow into the feed channel 15 only by the atmospheric pressure or gravity applied to the liquid material. Accordingly, in order to transfer the liquid material of the storage chamber 22 to the reaction chamber 25 through the feed transfer channel 15, a negative pressure enough to suck the liquid substance into the storage chamber 22 and the feed transfer channel 15. That is, a vacuum must be formed.

To this end, the liquid flow plate 10 of the present invention includes a discharge channel 41 and an eluate recovery channel connected to the reaction chamber 25, the mixing chamber 27, the eluent chamber 28 or the second reaction chamber 25a. 42, the air suction channel 43 or the liquid material suction channel 45 may be provided with a pump means for forming a pressure or a negative pressure. The pump means is for sucking or drawing air and is a conventional air pump or syringe.

As described above, the liquid flow plate 10 of the present invention may include a plurality of storage chambers 22 to store various types of liquid materials. In addition, the target material may be separated by mixing liquid materials or dispersing magnetic particles using the reaction chamber 25 and the mixing chamber 27 connected to the storage chamber 22. For this purpose, a predetermined magnetic application means may be installed in close proximity to the reaction chamber 25 of the liquid flow plate 10. Reference numeral 18 is an installation unit for installing such a magnetic application means. In addition, the installation unit 18 may be provided with devices for promoting a reaction, such as a heater or an ultrasonic generator.

According to the present invention, the magnetic particles for separating the target material are stored in the storage chamber 22 in a dispersed state in a solution. The magnetic particles stored in the storage chamber 22 are supplied to the reaction chamber 25 and react with the sample material to attach the target material. Subsequently, the magnetic particles to which the target substance is attached are fixed to the inner surface of the reaction chamber 25 by the action of the magnetic application means. The remaining solution except for magnetic particles is discharged to the outside through the discharge channel 41 connected to the mixing chamber 27.

The magnetic material fixed to the reaction chamber 25 falls off the wall of the reaction chamber 25 as the magnetic application means retreats and reacts with the reagent solution supplied from the storage chamber 22 again. The solution solution extracted from the sample may be extracted through the eluent chamber 28 connected to the reaction chamber 25 and the eluate recovery channel 42 connected to the eluent chamber 28.

In addition, according to another embodiment of the present invention, a second reaction, for example, DNA amplification, may be further performed using the second reaction chamber 25a connected to the reaction chamber 25. To this end, a heater for polymerase chain reaction (PCR) may be installed in the second reaction chamber 25a. In addition, an observation window 29 for observing the amplification result may be formed in the second reaction chamber 25a.

The liquid flow plate 10 according to the present invention may extract biological materials such as nucleic acids or proteins from various biological samples or amplify the extracted materials.

In addition, the liquid flow plate 10 of the present invention does not use consumables such as a tubular container, a seal or a tip because the tubular container or the seal or the tip performs all the functions performed in one plate.

In addition, the conventional target material extraction device required a complex and large scale device such as a tip feeder and a liquid handler for the injection, mixing and discharge of the liquid material, but using the liquid flow plate 10 of the present invention The structure of can be very simple.

Hereinafter, an embodiment of a target material extraction device using the liquid flow device according to the present invention will be described.

As shown in Figure 7 and 8, the target material extracting apparatus 200 of the present invention is largely coupled to the main body 201 and the upper surface of the main body 201 is mounted the liquid flow plate 10 according to the present invention It is composed of a cover 203 provided with a pump means 240 that is provided and coupled to the injection channel opening and closing means 220 and the discharge channel (41).

The upper surface of the main body 201 is provided with a mounting portion 205 for mounting the liquid flow plate 10. The mounting portion 205 consists of a groove sized to accommodate one liquid flow plate 10. And the lower end of the mounting portion 205 is provided with a magnetic application means 290 for fixing the magnetic particles on the inner surface of the reaction chamber (25). For example, the magnetic application means 290 is made of a permanent magnet installed so as to be movable up and down or made of an electromagnet equipped with a switch.

In addition, the cover 203 has injection channel opening and closing means 220 for selectively opening and closing a plurality of injection channels 13 formed on the upper surface of the liquid flow plate 10, and discharge formed on the upper surface of the liquid flow plate 10. The pump means 240 is coupled to the channel 41 to form a vacuum in the reaction chamber 25 or to discharge the liquid material. Preferably, the injection channel opening and closing means 220 and the pump means 240 interlock with each other to supply the liquid material of the storage chamber 22 to the reaction chamber 25 or to mix the liquid material inside the reaction chamber 25 or The liquid material in the reaction chamber 25 and the mixing chamber 27 is discharged to the outside.

More specifically, the injection channel opening and closing means 220 includes a plurality of valve bodies installed corresponding to the plurality of injection channels 13 and an actuator for operating the valve bodies. For example, the actuator may move the valve body up and down to open or close the injection channel 13. The pump means 240 is composed of a connection nozzle and a vacuum pump (or syringe) connected to the discharge channel (41).

In addition, the target material extraction apparatus 200 of the present invention is provided with control means (not shown) for controlling the injection channel opening and closing means 220 and the pump means 240. The control means operates the pump means 240 with any one of the plurality of injection channels 13 open. In addition, the target material extracting apparatus 200 of the present invention may be further provided with a control unit for inputting the data required for the control means and a display for showing the processing process. The control means is electrically connected to the actuators of the injection channel opening and closing means 220 and the pump means 240. In addition, the target material extracting apparatus 200 may be further provided with a heater, a laser or an ultrasonic device to promote the reaction of the liquid material.

Therefore, the target material extracting apparatus 200 according to the present invention supplies or reacts the liquid material stored in the storage chamber 22 to the reaction chamber 25 by using the injection channel opening and closing means 220 and the pump means 240. Two or more materials transferred to the chamber 25 are mixed and reacted. And by using the magnetic application means 290 to fix the magnetic particles in combination with the target material on the inner wall of the reaction chamber 25 it is possible to perform the process of extracting the biological material smoothly.

And, Figure 8 is another embodiment of the target material extracting apparatus 200 of the present invention, the discharge channel 41, the air suction channel 43 and the eluate recovery channel 42 of the liquid flow plate 10 is a liquid flow The case is formed on the side of the plate 10. Therefore, in the present embodiment, except for the above-described pump means 240 is installed in the main body 201, the rest of the configuration is the same as the above-described embodiment.

7 and 8 is a target material extraction method for extracting a nucleic acid from a biological material using the target material extraction apparatus 200 shown in Figure 9, as shown in Figure 9, the liquid material filling step (S10), the liquid flow plate It consists of a mounting step (S20), liquid material supply step (S30), liquid material mixing step (S40), magnetic particle fixing step (S50), liquid material discharge step (S60), eluent transfer and recovery step (S70).

The liquid material filling step (S10) is a step of injecting a predetermined liquid material into a plurality of storage chambers 22 provided in the liquid flow plate 10. The liquid substance includes various reagent solutions necessary for extracting biological substances such as sample solution, dispersion solution in which magnetic particles are injected, elution solution, cleaning solution, buffer solution and the like. This liquid material is injected through each injection channel 13 formed in the storage chamber 22. In the case of the sample solution, the pipette may be injected into the sample storage chamber through the injection hole 32. In addition, the injection channel 13 of the liquid flow plate 10 filled with the liquid material is sealed by a sealing means such as a tape or a sticker, and the injection hole 32 is sealed by using a rubber stopper 31. 10 shows a liquid flow plate 10 filled with liquid material in this manner.

The liquid flow plate mounting step (S20) is a step of seating the liquid flow plate 10 in which the liquid material is injected into the mounting portion 205 provided in the main body 201 and closing the cover 203. As shown in FIGS. 7 and 8, the main body 201 may be provided with one mounting unit 205, but a plurality of mounting unit 205 may be installed to install a plurality of liquid flow plates 10. When the liquid flow plate 10 is mounted and the cover 203 is covered, a pump corresponding to each of the discharge channel 41, the eluate recovery channel 42 and the air suction channel 43 of the liquid flow plate 10 is provided. The means 240 are in close contact with each other, and the valve bodies of the injection channel opening and closing means 220 are coupled to the corresponding injection channels 13, respectively. At this time, the tape or sticker sealing the injection channel 13 is removed in advance.

In this way, the extraction process is performed while the liquid flow plate 10 is mounted and the lid 203 is closed. This extraction process is a liquid material supply step (S30) for supplying the liquid material stored in the plurality of storage chambers 22 to the reaction chamber 25, and mixing two or more solutions supplied from the reaction chamber 13 or magnetic particles Liquid material mixing step for dispersing solid particles such as (S40), magnetic particle fixing step (S50) for fixing the dispersed magnetic particles to the reaction chamber 25 to separate the target material, and the reaction chamber (25) Liquid material discharge step (S60) for discharging the remaining solution to the outside and the eluent transfer and recovery step for transferring the eluate in the reaction chamber 25 to the eluent chamber 28 or to the outside (S70) ) This extraction process is performed by controlling the injection channel opening and closing means 220 and the pump means 240.

First, the liquid material supply step (S30) is to operate the pump means 240 is connected to the discharge channel 41, to form a vacuum or low pressure in the reaction chamber 25 and the mixing chamber 27 and In addition, by opening one or more valve body of the injection channel 13 of the injection channel opening and closing means 220, the corresponding storage chamber 22 is in communication with the outside air to suck the air of the storage chamber 22 Allow the interior to be at atmospheric pressure. Then, the liquid substance at atmospheric pressure stored in the storage chamber 22 is supplied to the reaction chamber 25 along the corresponding feed transfer channel 15 by the vacuum or low pressure formed in the reaction chamber 25 (see FIG. 11).

Then, the liquid material mixing step (S40), by operating the pump means 240 connected to the air suction channel 43 to suck the liquid material in the reaction chamber 25 into the reaction chamber 25 and Repeating the step of discharging the liquid material in the mixing chamber 27 to the reaction chamber 25. For example, by operating the pump means 240 to form a low pressure inside the mixing chamber 27 and the injection channel opening and closing means 220 to operate the injection channel of the storage chamber 22 is empty liquid material When opening 13, the liquid substance inside the reaction chamber 25 is transferred to the mixing chamber 27 through the feed transfer channel 16 by the vacuum of the mixing chamber 27 (see Fig. 12).

On the contrary, the pump means 240 operates to inject air into the mixing chamber 27 and the injection channel opening and closing means 220 to operate the injection channel 13 of the storage chamber 22 in which the liquid material is empty. ), The liquid material in the mixing chamber 27 is transferred to the reaction chamber 25 through the feed transfer channel 16 (see FIG. 13). The target material may be attached to the surface of the magnetic particles by mixing two or more liquid materials evenly or dispersing the magnetic particles contained in the liquid material without rotating or shaking the plate 10 itself.

Then, the magnetic particle fixing step (S50) is made by the action of the magnetic application means 290 not shown. For example, when electricity is applied to the electromagnet or the permanent magnet 290 is approached to the reaction chamber 25, the magnetic particles P dispersed in the reaction chamber 25 are applied to the magnetic field of the magnetic application means 290. By being agglomerated and fixed to the inner wall of the reaction chamber 25 (see FIG. 14).

Subsequently, the liquid material discharge step (S60) is performed while the magnetic particles P are fixed to the inner wall of the reaction chamber 25. For example, by operating the pump means 240 connected to the discharge channel 41 to apply a vacuum to the discharge pump 241 while using the injection channel opening and closing means 220, the storage chamber empty of the liquid material ( Opening the injection channel 13 of the 22, the reaction chamber 25 and the mixing chamber 27 connected to the storage chamber 22 is in the atmospheric pressure state to discharge the residual liquid material inside the reaction chamber 25 to the outside (See FIG. 15)

And the eluate transfer and recovery step (S70), the process of transferring the eluent in the reaction chamber 25 to the eluent chamber 28, and the process of discharging the eluent in the eluent chamber 28 to the outside This eluate transfer and recovery process is performed by operating the pump means 240 connected to the eluate recovery channel 42 to form a vacuum in the recovery channel 42 and the liquid phase using the injection channel opening and closing means 220. This is achieved by opening the injection channel 13 of the storage chamber 22 in which the material is empty. Then, the reaction chamber 25 connected to the storage chamber 22 is at atmospheric pressure so that the eluate in the reaction chamber 25 is transferred to the eluent chamber 28, and when the pump means 240 continues to operate, the eluent The eluate in the chamber 28 is discharged to the outside through the eluate recovery channel 42. The eluent transfer step and the recovery step S70 may be performed in one continuous process, but the step of transferring the eluate and the recovery of the eluate may be performed in separate processes. In particular, the eluate recovery step may be performed using a pipette as in the prior art.

In the above, the steps of constituting the target material extraction method have been described based on important steps. However, these steps may be changed according to a sample or a target material. Therefore, the steps according to the present invention may be repeatedly performed or the order of these steps may be changed depending on the type of target material or reagent. In addition, the present invention may extract nucleic acids using a plurality of liquid flow plates, but in this case, the basic process may be equally applied.

Meanwhile, as described above, the liquid flow plate according to the present invention may stack a plurality of unit liquid flow plates. That is, the liquid flow plate of the present invention is to control the flow of the liquid material through the opening and closing of the injection channel instead of controlling the valve installed on the transfer channel, it is possible to stack a plurality of liquid flow plate. Therefore, hereinafter, a liquid flow block formed by stacking a plurality of liquid flow plates according to the present invention, an apparatus for extracting a target substance using the same, and a method for extracting the target substance will be described.

First, FIGS. 17 and 18 show a liquid flow block 300 formed by stacking a unit liquid flow plate 100 and a plurality of unit liquid flow plates 100 as another embodiment of the liquid flow apparatus according to the present invention.

First, Figure 17 is a perspective view showing a unit liquid flow plate 100 according to the invention. The unit liquid flow plate 100 according to the present invention has the same structure as the liquid flow plate 10 described above except that the injection channel 113 completely passes through the liquid flow plate 10.

As shown, the unit liquid flow plate 100 has a flat plate shape, and a plurality of storage chambers 22 are formed at an upper end thereof. An injection channel 113 corresponding to each of the storage chambers 22 is formed in the plurality of storage chambers 22. The injection channel 113 is connected to the upper end of the storage chamber 22 and a narrow neck is provided between the injection channel 13 and the storage chamber 22.

On the other hand, the upper surface of the unit liquid flow plate 100 may be further provided with a sample storage chamber for injecting a sample solution using a pipette. In addition, an injection hole 32 for injecting a sample using a pipette is formed at an upper portion of the sample storage chamber. And the injection hole 32 is provided with a rubber stopper 31 for preventing the inflow of air and foreign matter. In addition, a recovery port 34 capable of recovering the eluate by using a pipette may be further formed on the eluent chamber 28. In addition, the recovery port 34 is also provided with a rubber stopper 33 to prevent the inflow of air and foreign matter and insert the injection needle to extract the eluate.

As shown in FIG. 18, the liquid flow block 300 according to the present invention is formed by stacking a plurality of unit liquid flow plates 100 described above. For example, the liquid flow block 300 is formed by stacking the unit liquid flow plates 100: 100A, 100B, 100C ..... 100H in a vertical direction in a horizontal direction (left and right directions). In this case, the liquid flow block 300 is formed such that the injection channel 113 of the neighboring unit liquid flow plate 100 is positioned on the same line. In addition, an O-ring (not shown) is inserted between neighboring injection channels 113 to prevent leakage of the liquid material. In addition, the plurality of unit liquid flow plates 100 are tightly coupled by using a band, a cartridge, or a tray, so that the plurality of injection channels 113 are arranged in a line and extend from one side to the other side of the liquid flow block 300. The injection channel 313 is formed.

As such, in the liquid flow block 300 according to the present invention, a plurality of injection channels 313: 313a, 313b,... 313g are formed at both sides, and a plurality of eluents are provided at the front of the liquid flow block 300. Recovery channels 42: 42A, 42B ... 42H are formed, and a plurality of discharge channels 41: 41A, 41B ... 41H and air suction channels 43: 43A, 43B ... 43H are formed on the rear side. do. In addition, a plurality of injection holes 32 for injecting a sample solution and a plurality of recovery holes 34 for recovering the eluate are formed on the upper surface of the liquid flow block 300. In addition, the injection hole 32 and the recovery port 34 are provided with rubber stoppers 31 and 33, respectively.

As such, the liquid flow block 300 according to the present invention is provided with a plurality of reaction chambers 25: 25A, 25B ... 25H therein, so that the target substance can be extracted from a large amount of biological sample at a time. . In addition, the plurality of storage chambers 22: 22A, 22B, 22C ... 22H are connected to one injection channel 313 in the liquid flow block 300 of the present invention. 22A, 22B, 22C ... 22H) can automatically fill the same liquid material. That is, in the related art, a researcher needs to fill a plurality of reaction vessels with the same liquid material in a predetermined amount using a pipette, but it takes a long time and is very troublesome, but the liquid flow block 300 of the present invention has a plurality of storage chambers 22. : 22A, 22B, 22C ... 22H) are connected to each other easily to automate the quantitative filling of the same liquid material. The liquid filling method and the nucleic acid extraction process of the liquid material is achieved by the target material extraction apparatus described below.

13 is an explanatory view showing the internal configuration of the liquid flow block 30 according to the present invention. As shown, a plurality of unit liquid flow plate 100 is stacked, and a plurality of injection channels 113a: 113Aa, 113Ba, 113Ca.... 113Ha are connected to the left and right to form one injection channel ( 313a), a plurality of injection channel channels 113b: 113Ab, 113Bb, 113Cb ..... 113Hb are connected to the left and right to form one injection channel 313b, and a plurality of injection channels 113g: 113Ag, 113Bg, 113Cg ..... 113Hg) is connected to the left and right to form one injection channel (313g).

Each of the injection channels 313a, 313b... 313g is connected with a plurality of storage chambers 22 for storing the same substance in the same amount. For example, a plurality of storage chambers 22Aa, 22Ba, 22Ca .... 22Ha are connected in a line to the injection channel 313a, and a plurality of storage chambers 22Ag, 22Bg, 22Cg ... 22Hg are connected to the injection channel 313g. Are connected in line.

Therefore, when the sample solution is injected into the injection channel 313a, a predetermined amount of the sample solution is injected into the plurality of storage chambers 22Aa, 22Ba, 22Ca .... 22Ha, and the solution is injected into the injection channel 313g. The chambers 22Ag, 22Bg, 22Cg ... 22Hg are filled with a certain amount of solution. As described above, the method of quantitatively filling the plurality of storage chambers 22 with the liquid material through the injection channel 313 will be described below.

20 to 21 show a preferred embodiment of the target material extraction apparatus 500 using the liquid flow block 300 according to the present invention.

As shown, the target material extraction apparatus 500 using the liquid flow block 300 according to the present invention, the main body 502 and the upper portion of the main body 502 for mounting the liquid flow block 300 of the present invention It consists of a cover 503 for covering the opening.

The main body 502 has a mounting portion 505 for mounting the liquid flow block 300 and an injection channel 313, a discharge channel 41, and an eluate recovery of the liquid flow block 300 mounted on the mall mounting portion 505. The pump unit 240 is in close contact with the channel 42 and the air suction channel 43, and an operating unit 510 is provided to perform an operation for injecting and supplying, transporting, or discharging the liquid material.

The mounting part 505 is formed of a groove having a predetermined depth to which the lower end of the liquid flow block 300 is inserted and fixed at a predetermined depth. The operation unit 510 is configured to be in close contact with each surface of the liquid flow block 300 mounted on the mounting unit 505, the left operation is installed on the left and right around the mounting unit 505 The unit 512 and the right operation unit 514, the front operation unit 516 and the rear operation unit 518 is installed in the front and rear.

The left operating part 512 and the right operating part 414 are installed to be horizontally movable in the left and right directions, and the front operating part 516 and the rear operating part 518 are installed to be horizontally movable in the front-rear direction. To this end, an operation means for operating the operation unit 510 is provided inside the main body 502. For example, the actuation means comprises a motor, a power transmission means and a guide means. Since such a configuration is well known in the art, a detailed description thereof will be omitted.

In this case, the mounting unit 505 is provided with a sensor for detecting whether the liquid flow block 300 is mounted, the operation unit 510 is activated when the mounting of the liquid flow block 300 by the sensor. For example, when the liquid flow block 300 is mounted to the mounting portion 505, the left operation portion 512 and the right operation portion 514 move to the left and right sides, and the front operation portion 516 and the rear operation. The unit 518 is horizontally moved to the front and rear sides, respectively, so that the pump means 240 is in close contact with the channel formed on each side of the liquid flow block 300.

In addition, a magnetic applying means 590 is provided below the mounting portion 505 of the main body 502. The magnetic application means 590 is installed at a position corresponding to the installation portion 18 formed in the liquid flow block 300. The magnetic application means 590 is composed of a permanent magnet installed to be movable up and down or an electromagnet equipped with a switch. As shown in FIG. 13, a display unit 506 and an input unit 507 are provided at one side of the main body 502.

Next, referring to FIG. 22, a liquid material and air supply means 420 is installed in the left operating part 512 of the operating part 510 according to the present invention, and a liquid material and air suction is installed in the right operating part 514. Means 440 are installed. In addition, the liquid transfer and discharge means 460 is installed in the rear operation part 518, and an eluate transfer and recovery means 480 is installed in the front operation part 516. The liquid material and air supply means 420 and the liquid material and air suction means 440 performs the same function as the injection channel opening and closing means 220 for controlling the injection channel 13 in the above-described embodiment, It further performs the function for automatic injection. The liquid material transfer and discharge means 460 and the eluent transfer and recovery means 480 perform the same functions as the pump means 240. The liquid material and air supply means 420, the liquid material and air suction means 440, and the liquid material transfer and discharge means 460 and the eluate transfer and recovery means 480 are installed at the injection channel 13. However, since the discharge channel 41, the air suction channel 43, and the eluate recovery channel 42 vary depending on the positions, the present invention is not limited thereto. However, since the liquid material and the air supply means 420 and the liquid material and the air suction means 440 are respectively installed at both inlets of the injection channel 313, they are always located at both sides of the liquid flow block 300.

Referring again to FIG. 22, the liquid material and air supply means 420 is for injecting liquid material and air through a plurality of injection channels 313 formed on the left side of the liquid flow block 300. The liquid material and air suction means 440 is for sucking the liquid material and air through a plurality of injection channels 313 formed on the right side of the liquid flow block 300. That is, the liquid material and the air supply means 420 and the liquid material and the air suction means 440 is connected to each other to fill the liquid material in the storage chamber 22 through the injection channel 313 or the storage chamber 22 Inject air into the air to ensure atmospheric pressure. In addition, the liquid material and air supply means 420 and the liquid material and air suction means 440 selectively opens and closes a plurality of injection channels 313 to react the liquid material stored in the storage chamber 22 with the reaction chamber 25. Or to discharge the liquid material of the reaction chamber 25 to the outside.

As shown, the liquid material and air supply means 420, a plurality of main supply pipe 424 and the liquid material to be tightly coupled to the plurality of injection channels 313 formed in the liquid flow block 300, A plurality of liquid material tanks 422, a liquid material supply pipe 426 for supplying a liquid material by connecting the main supply pipe 424 and a liquid material tank 422, and a plurality of liquid material supply pipes 426. The liquid material control valve 428 is installed, the plurality of air supply pipes 436 connected to the plurality of main supply pipes 424 and the other end is in communication with the outside air, and are installed in the air supply pipes 436 to control the flow of air. A plurality of intermittent air supply control valve 438 is made up.

At this time, the front end of the main supply pipe 424 is made to pass through the left connecting portion 513 installed to be movable left and right. Accordingly, the main supply pipe 424 may be closely connected to the injection channel 313 by moving only the left connection part 513 from side to side without moving the left operating part 512 from side to side.

The liquid material and air suction means 440 includes a plurality of main suction pipes 444 closely coupled to a plurality of injection channels 313 formed on the other side of the liquid flow block 300, and the main suction pipes 444. A plurality of main pumps 445 installed to be connected to the injection channel 313 to form a vacuum in the injection channel 313, a liquid material drain pipe 446 connected to the main pump 445 to discharge liquid materials, and A plurality of main control valves 448 installed in the plurality of main suction pipes 444 to open and close the main suction pipes 444, an air discharge pipe 447 connected to the main pump 444 to discharge air, and the air It is configured to include an air discharge control valve 449 installed in the discharge pipe (447).

At this time, the front end of the main suction pipe 444 is made through the right connecting portion 515 installed to be movable left and right, and thus only the right connecting portion 515 without moving the right operating portion 514 from side to side. The main suction pipe 444 may be connected to the injection channel 313 by moving from side to side.

Subsequently, the liquid material transfer and discharge means 460 includes a plurality of liquid material discharge pipes 461 connected to the plurality of discharge channels 41 provided on the rear surface of the liquid flow block 300, and the plurality of liquid materials. A main liquid material discharge pipe 464 connected to the discharge pipe 461, a discharge pump 431 installed on the main liquid material discharge pipe 464, a liquid material drain pipe 466 connected to the discharge pump 431, and A liquid material discharge control valve 468 installed in the main liquid material discharge pipe 464 to control discharge of the liquid material, and a remaining liquid material storage tank 462 connected to the liquid material drain pipe 466. .

In addition, a plurality of air discharge pipes 463 connected to the plurality of air suction channels 143 provided on the rear surface of the liquid flow block 300, and a main air pipe discharge pipe 465 connected to the plurality of air discharge pipes 463. And a mixing pump 433 connected to the main air discharge pipe 465, an air exhaust pipe 467 connected to the mixing pump 433, and installed in the main air discharge pipe 465 to control the flow of air. It comprises an air discharge control valve 469. In the drawing, the liquid material discharge pipe 461 and the air discharge pipe 463 are shown to overlap, but in fact, they are separated up and down and formed separately.

The eluate transfer and recovery means 480 includes a plurality of eluate discharge pipes 482 connected to a plurality of eluate recovery channels 42 provided on the front surface of the liquid flow block 300, and the plurality of eluate discharge pipes 482. Eluent discharge control valve (488) installed in the main eluent discharge pipe 484, the eluent pump 432 connected to the main eluent discharge pipe (484), and the main eluent discharge pipe (484) to control the discharge of the eluate And an eluate recovery tube 489 connected to the eluate pump 432.

In addition, the target material extracting apparatus 500 according to the present invention is the liquid material and air supply means 420, the liquid material and air suction means 440, liquid material transfer and discharge means 460, eluent transfer and recovery Control means for controlling the various valves and pumps provided in the means 480 is further included. In addition, the control means is electrically connected to various sensors, operating means (motor) and the magnetic application means (490).

Therefore, the target material extracting apparatus 500 according to the present invention is the liquid material and air supply means 420 and the liquid material and air suction means 440, the liquid material and air supply means 420 and the liquid material transfer and discharge Means 460 or the liquid material and air supply means 420 and the eluent transfer and recovery means 480 to fill the liquid material in the storage chamber 22, the reaction of the liquid material stored in the storage chamber 22 The liquid is transferred to the chamber 25 and the liquid material is sucked and discharged between the reaction chamber 22 and the mixing chamber 27 to mix and discharge the liquid material. The magnetic application means 490 is controlled to fix the magnetic particles inside the reaction chamber 25 so that the nucleic acid material can be extracted from various biological samples.

Hereinafter, a method of extracting nucleic acid from a biological sample using a target material extraction device using a liquid flow block according to the present invention will be described.

As shown in Figure 23, the target material extraction method of the present invention, large liquid flow block mounting step (S100), liquid material filling step (S200), liquid material supply step (S300), liquid material mixing step (S400), Magnetic particle fixing step (S500), liquid material discharge step (S600), eluent transfer and recovery step (S700).

First, the liquid flow block mounting step (S100) is a step of mounting the liquid flow block 300 on the main body 502 of the target material extracting apparatus 500 illustrated in FIGS. 20 and 21. In this case, the liquid flow block 300 may be filled or not filled with a liquid material in each storage chamber 22, but the present invention will be described for the case where the liquid material is not filled. In addition, when the liquid flow block 300 is wrapped in a plastic wrap to prevent contamination, the liquid flow block 300 is mounted on the mounting portion 505 of the main body 502 while the plastic wrap is peeled off.

Then, the detection sensor (not shown) provided in the main body 502 detects this, and according to the control of the control means not shown, the left operation part 512 and the right operation part 514 or the front operation of the operation part 510 are performed. The unit 516 and the rear operation unit 518 are horizontally moved to closely contact each side of the liquid flow block 300. At this time, when the operation unit 51 is provided with the left connector 512 and the right connector 515 described above, the left operating unit 512 and the right operating unit 514 are not moved by horizontally moving these connectors. It may be in close contact with both sides of the liquid flow block 300 without.

Then, the plurality of main supply pipes 424 and the inlet suction pipe 444 provided in the left operating part 512 are connected to the injection channel 313 of the liquid flow block 300. And the plurality of liquid material discharge pipe (461) provided in the rear operation unit 518 is connected to the plurality of discharge channels 41 provided on the rear of the liquid flow block 300, a plurality of air discharge pipe (463) It is connected to a plurality of air intake channels 43 provided on the rear of the liquid flow block 300. In addition, the plurality of eluate discharge pipes 482 provided in the front operation part 516 are connected to the plurality of eluate recovery channels 42 provided on the front surface of the liquid flow block 300, respectively. In this case, a connection nozzle is provided at the front end of the main supply pipe 424, the intake suction pipe 444, the liquid material discharge pipe 461, the air discharge pipe 463, and the eluate discharge pipe 482 to discharge the channel of the liquid flow block 300. 41, the eluent recovery channel 42 and the air suction channel 43 is in close contact with each other to prevent the leakage of the liquid material (see Fig. 24).

Subsequently, the liquid material filling step (S200) is a process of filling a predetermined amount of a predetermined liquid material into the plurality of storage chambers 22, and is further divided into a liquid material injection step (S210) and an air injection step (S220). The liquid material injection step (S210) and the air injection step (S220) of the liquid material and air supply means 420 of the left operating part 512 and the liquid material and air suction means 440 of the right operation part 514 It is achieved by interlocking.

First, the liquid material injection step (S210) operates a plurality of main pumps 445 provided on the right operating part 514 to form a vacuum in the main suction pipe 444 and is provided on the left operating part 512. Close all of the plurality of air control valves 438, and open all of the plurality of liquid material control valves 428 connected to the liquid material storage tank 422, and thus, the injection channel 313 of the liquid flow block 300. The liquid material of the liquid material storage tank 422 is injected into the injection channel 313 by the low pressure formed in the).

Then, the liquid material transferred through the injection channel 313 is sequentially filled in the plurality of storage chambers 22 connected to the injection channel 313 so that the liquid material is filled in each of the storage chambers 22 and the injection channel 313. Will be filled. In this process, some of the liquid material may be introduced into the liquid material recovery tank 443 through the main drain pipe 446 connected to the main pump 445 (see FIG. 25).

Subsequently, the air injection step (S220) discharges a portion of the liquid material filled in the injection channel 313 and a portion of the liquid material stored in the storage chamber 22 to the outside of the liquid material injected into each storage chamber 22. As a process of uniformly adjusting the water level, that is, the volume of the liquid substance, the main pump 445 provided by the right operating part 514 is operated to form a vacuum in the main suction pipe 444 and at the same time the left operating part 512. Close all of the plurality of liquid substance control valve 428 provided in the air, and open all the air control valve 438 of the plurality of air supply pipe 436 connected to the outside air to inject external air into the injection channel 313 Be sure to

Then, since the liquid material of the injection channel 313 and a part of the liquid material stored in the storage chamber 22 are discharged to the outside, the level of each storage chamber 22 is constant. That is, the liquid material of the storage chamber 22 is infused with the injection channel 313 until the suction force formed in the injection channel 313 and the surface tension and gravity applied to the liquid material in the storage chamber 22 are balanced. Since it is discharged through), the level of the liquid material in each storage chamber 22 is constant. In this case, the liquid material discharged from the injection channel 313 may be removed through the main drain pipe 446 or may be recovered to the liquid material recovery tank 443. (See FIG. 26)

As described above, the present invention can omit a conventional pipette operation because the plurality of storage chambers 22 can be automatically filled with a plurality of liquid substances. In addition, since the liquid material is supplied after the liquid flow block 300 is mounted on the target material extraction device, not only the contamination of the liquid material can be prevented, but also a separate pretreatment device can be omitted, thereby making it compact. In addition, if necessary, the pipette may be injected into the sample storage chamber through the injection hole 32 formed on the upper surface of the liquid flow block 300.

Subsequently, the liquid material supply step (S300), liquid material mixing step (S400), magnetic particle fixing step (S500), liquid material discharge step (S600), eluent transfer and recovery step (S700) is to extract the nucleic acid from the biological sample Nucleic acid extraction process for. This nucleic acid extraction process is similar to the extraction process of the liquid flow plate 10 described above. In addition, the steps of the extraction process may be performed repeatedly by changing the order of the steps or connecting one or two steps according to the target material and the type of reagent.

First, the liquid phase material supplying step S300 is a process of supplying the liquid phase material stored in the plurality of storage chambers 22 connected to the specific injection channel 313 to the plurality of reaction chambers 25. That is, by operating the discharge pump 431 provided in the rear operation unit 518 in the reaction chamber 25 and the mixing chamber 27 provided in the liquid flow block 300 connected to the liquid material discharge pipe 461. The air control valve 438 of the air supply pipe 436 which forms a vacuum pressure and is connected to a specific injection channel 313, that is, an injection channel 313 into which a liquid substance to be supplied to the reaction chamber 25 is injected. When the outside air is introduced into each of the storage chambers 22 through the injection channel 313, the storage chamber 22 is at atmospheric pressure, and the liquid substance inside the storage chamber 22 reacts. The suction force of the chamber 25 is transferred to the reaction chamber 25 through the supply transfer channel 115. In this case, in some cases, the liquid material may be transferred to the mixing chamber 27 through the mixing transfer channel 116. Subsequently, when all the liquid material of the storage chamber 22 is supplied, the open supply control valve 438 is closed and the discharge pump 431 is operated in the opposite direction to store some liquid material introduced into the mixing chamber 27 again. The liquid material supply step is completed by sending it to the chamber 22. At this time, all air control valves 438, all liquid material control valves 428, and all main control valves 448 except the air control valve 438 connected to the specific injection channel 313 remain closed. . By repeating this process, the liquid materials stored in the plurality of storage chambers 22 may be sequentially transferred and reacted (see FIG. 27).

The liquid material mixing step (S400) is a process for mixing two or more liquid materials or samples and magnetic particles supplied to the reaction chamber 25, in particular, from the reaction chamber 25 to the mixing chamber 27 or mixing. It is a process of repeatedly transferring the liquid material from the chamber 27 to the reaction chamber 25. To this end, the suction step of operating the mixing pump 433 provided in the rear operation unit 518 to form a vacuum pressure in the mixing chamber 27 and the mixing pump 433 in the opposite direction to operate the mixing The discharging step of forming pressure in the chamber 27 is repeatedly performed.

For example, the suction step is to form a vacuum pressure in the mixing chamber 27 and at the same time the injection of the liquid material in the storage chamber 22 is removed in a specific injection channel 313, that is, the liquid material supply step (S300) The air control valve 438 of the air supply pipe 436 connected to the channel 313 is opened to allow external air to flow into the reaction chamber 25 through the injection channel 313 and the storage chamber 22. Then, the reaction chamber 25 is at atmospheric pressure, so that the liquid substance inside the reaction chamber 25 is sucked into the mixing chamber 27 along the feed transfer channel 16 by the suction force of the mixing chamber 27. do. On the contrary, when the liquid material is filled in the mixing chamber 27, the mixing pump 433 provided in the rear operation part 518 is operated in reverse to blow air into the mixing chamber 27, and at the same time, a specific injection channel 313 ), Ie, when the air control valve 438 of the air supply pipe 436 is connected to the injection channel 313 in which all the liquid material of the storage chamber 22 is removed in the liquid material supplying step (S300), The liquid substance inside the mixing chamber 27 is discharged to the reaction chamber 25 in the atmospheric pressure along the supply transfer channel 16. By repeating the suction and discharge steps, the sample and the reagent or the target material and the magnetic particles contained in the liquid material are sufficiently mixed. At this time, all air control valves 438, all liquid material control valves 428, and all main control valves 448 except the air control valve 438 connected to the specific injection channel 313 are closed.

The magnetic particle fixing step (S500) is a process of aggregating and fixing the magnetic particles on the inner wall of the reaction chamber 25, and mainly discharges the cleaning solution for cleaning the remaining reagent solution and foreign matter reacted with the sample or recover the eluate When desired, the magnetic particles dispersed in the reaction chamber 25 are fixed to the inner wall of the reaction chamber 25. Magnetic particle fixing step (S500) is made by the magnetic application means 490 provided in the body 502. For example, when electricity is applied to the electromagnet of the magnetic application means 490 or the permanent magnet is approached to the reaction chamber 25, the dispersed magnetic particles in the reaction chamber 25 may be separated from the magnetic application means 490. Aggregated by the magnetic field and fixed to the inner wall of the reaction chamber 25. In this manner, the remaining liquid substance may be discharged or the eluate is recovered while the magnetic particles are fixed to the inner wall surface of the reaction chamber 25 (see FIG. 28).

The liquid material discharging step (S600) is a process of discharging the remaining liquid material (reagent or cleaning liquid, etc.) except for the magnetic particles in a state where the magnetic particles are fixed to the reaction chamber 25, and are provided in the rear operation part 518. By operating the discharge pump 431 to form a vacuum and the air control valve 438 of the air supply pipe 436 which is connected to a specific injection channel 313, that is, the injection channel 313 from which the liquid substance is removed. When opened, external air flows into the reaction chamber 25 through the injection channel 313 and the storage chamber 22, and the reaction chamber 25 is at atmospheric pressure. Then, the liquid material in the reaction chamber 25 is sucked into the discharge pump 431 through the mixing chamber 27 and the discharge channel 41 by the suction force of the discharge pump 431. The liquid material sucked into the discharge pump 431 is discharged to the remaining liquid material storage tank 462 through the liquid material drain pipe 466 (see FIG. 29).

Subsequently, the eluate transfer and recovery step (S700) is a process of transferring or recovering an eluate containing a target substance except the magnetic particles in a state in which magnetic particles are fixed in the reaction chamber 25. Conveying step and recovering the eluate in the eluate chamber 28. The eluate transfer step is to operate the eluent pump 432 provided in the front operation unit 516 to form a vacuum in the eluent chamber 28 provided in the liquid flow block 300 and to remove the liquid material When the air control valve 438 of the air supply pipe 436 connected to the injection channel 313 is opened, the reaction chamber 25 connected to the outside air is at atmospheric pressure and the eluate chamber 28 is at low pressure. The eluate in the reaction chamber 25 is transferred to the eluate chamber 28 through the eluate transfer channel 17. The eluate recovery step is a step of recovering the eluate in the eluate chamber 28 to an external container through an eluate drain tube 487 connected to the eluate pump 432. On the other hand, the eluate recovery step may be made through a recovery port 34 formed on the upper surface of the liquid flow block 300 using a separate pipette without using the eluent pump 432 (see Fig. 30).

As described above, the target material extracting apparatus 500 using the liquid flow block 300 according to the present invention is necessarily in the liquid material filling step (S200) and nucleic acid extraction to automatically fill the liquid material in the storage chamber 22 By supplying the necessary liquid material, mixing, discharging step and magnetic particle fixing step in one liquid flow block 300, it is possible to quickly process a large amount of bio information.

In addition, the target material extracting apparatus 500 of the present invention can reduce the size of the target material extracting apparatus because it uses a liquid flow device that can perform various functions unlike the conventional tubular container or multi-well plate kit. It can do the pretreatment process quickly and easily by automatically dosing various kinds of liquid materials, and can reduce the use of consumables by not using tips or seals to fix or move magnetic particles. Alternatively, the transfer device or the loading means for moving the pipette, in which the magnetic particles are fixed, up, down, left, and right, is unnecessary, thereby simplifying the structure of the target material extraction device.

Hereinafter, a liquid dosing device 700 according to the present invention will be described with reference to FIG. 31. The liquid metering device 700 according to the present invention includes an injection channel 313 connected to a plurality of storage chambers 22 and upper ends of the plurality of storage chambers 22 and the plurality of storage chambers 22. Liquid quantitative flow block 710 consisting of a discharge channel 41 connected to the lower end, respectively, liquid material and air supply means 720 and the liquid quantitative flow is installed on one side of the liquid quantitative flow block 710 The liquid material and the air suction means 740 is installed on the other side of the block 710. And the lower end of the liquid metering flow block 710 is provided with a container or a multi well plate kit 750 for filling the liquid material.

The liquid material and air supply means 720 is installed on the left side of the liquid metering flow block 710 to inject liquid material and air through a plurality of injection channels 313, and the liquid material and air suction The means 740 is installed on the right side of the liquid metering flow block 710 to suck liquid material and air through a plurality of injection channels 313. That is, the liquid material and air supply means 720 and the liquid material and air suction means 740 are connected to each other to supply the liquid material to the storage chamber 22 through the injection channel 313 or to the storage chamber 22. Air is injected into the inside to discharge the liquid substance stored in the storage chamber 22 to the container or the multi well plate kit 750 through the discharge channel 41.

More specifically, the liquid material and air supply means 720, the plurality of main supply pipe 724 and the liquid supply material for tightly coupled to the plurality of injection channels 313 formed in the liquid metering flow block 710, A plurality of liquid material tanks 722, a liquid material supply pipe 726 for supplying a liquid material by connecting the main supply pipe 724 and a liquid material tank 722, and installed in the plurality of liquid material supply pipes 726. Liquid material control valve 728, a plurality of air supply pipe 736 connected to the plurality of main supply pipe 724 and the other end is in communication with the outside, and is installed in the air supply pipe 736 to regulate the flow of air It consists of a plurality of air supply control valve 738.

The liquid material and air suction means 740, a plurality of main suction pipe 744 and the main suction pipe 444 closely coupled to a plurality of injection channels 313 formed on the other side of the liquid metering flow block 710, A plurality of main pumps 745 installed to be connected to the injection channel 313 to form a vacuum in the injection channel 313, a liquid material drain pipe 746 connected to the main pump 745 to discharge liquid materials; A plurality of main control valves 748 installed at the plurality of main suction pipes 744 to open and close the main suction pipes 744, an air discharge pipe 747 connected to the main pump 744 to discharge air, and the It is configured to include an air discharge control valve 749 installed in the air discharge pipe 747. In addition, the liquid material drain pipe 746 may be further provided with a liquid material storage tank 743.

The liquid substance and air supplying means 720 and the liquid substance and air suctioning means 740 are interlocked with each other through a controller (not shown).

Therefore, by using the quantitative supply device 700 of the present invention, it is possible to quantitatively supply various kinds of liquid materials to the container or multi well plate kit 750. In the liquid quantitative supply method of the present invention, as shown in FIG. 32, a plurality of injection channels 313 formed in the liquid quantitative flow block 710 and a plurality of storage chambers 22 connected to the injection channel 313 are provided. And a liquid material filling step of injecting a liquid material into S210; An air suction step (S220) for sucking air in the injection channel 313 to equalize the level of the liquid material injected into the plurality of storage chambers 22; It consists of a liquid material discharge step (S230) for discharging the liquid material stored in the storage chamber 22 to the outside.

First, the liquid material filling step (S210), by operating the main suction pump 745 connected to the injection channel 313 of the liquid metering flow block 710, the main suction passage 744 and the injection channel ( A low pressure is formed in the inside of the 313 and the air control valve 738 installed in the air supply pipe 736 connected to the injection channel 313 of the liquid metering flow block 710 is closed. Open the liquid material control valve 728 installed in the liquid material supply pipe 726 connected to the 313, the liquid material of the liquid material tank 722 flows into the injection channel 313 through the main supply pipe 724 The liquid material injected into the injection channel 313 is sequentially filled in the storage chamber 22 connected to the injection channel 313. The liquid material filling step is performed until all of the liquid material is filled in the plurality of storage chambers 22 and the liquid material is partially or partially filled in the injection channel 313. In addition, the liquid material flowing into the main suction pipe 744 is discharged through the main drain pipe 746 (see FIG. 33).

Then, the air suction step (S220), by operating the main suction pump 745 connected to the injection channel 313 of the liquid metering flow block 710, the main suction passage 744 and the injection channel ( In addition to forming a low pressure inside the 313, the air control valve 738 is opened in a state where the liquid substance control valve 728 is closed, so that air from the outside air is injected through the main supply pipe 724. Is injected into the channel 313 to be discharged to the outside through the main suction pipe 744 and the main drain pipe 746. Then, a low pressure is formed at the upper bottleneck of the storage chamber 22 by the flow of air flowing along the injection channel 313, so that a certain amount of liquid material is stored in the injection channel ( 313) is discharged to the outside. In this case, since the size and shape of the plurality of storage chambers 22 connected to the injection channel 313 are the same, the amount of the liquid substance discharged to the injection channel 313 is the same. Therefore, the liquid level of the liquid material in the storage chamber 22 is equal, so that the same amount of liquid material is stored in each storage chamber 22. Therefore, the air suction step (S220) is performed until the level of the liquid material stored in the plurality of storage chambers 22 becomes the same (see Fig. 34).

Subsequently, the liquid material discharging step (S230) is to discharge the liquid material stored in the storage chamber 22 to supply the quantitatively to the container or the multi well plate kit 750, and the injection channel of the liquid quantitative flow block 710. By operating the main suction pump 745 connected to the 313 to form a high pressure in the main suction passage 744 and the injection channel 313, the liquid material control valve 728 and air When the control valve 738 is closed, high pressure is formed in the storage chamber 22 so that the liquid substance stored in the storage chamber 22 is discharged to the container or the multi well plate kit 750 through the discharge channel 41. do.

As described above, the liquid quantitative supply device and the liquid quantitative supply method using the same according to the present invention are conventional pipettes because a plurality of injection channels 313 and a plurality of storage chambers 22 connected to the plurality of injection channels 313 are connected. It is possible to supply a quantitative amount of liquid substance without using. In addition, since it is possible to supply the liquid material in a quantitative manner to a plurality of containers, it is very convenient when quantitatively supplying a large amount of liquid material. In addition, the liquid metering device of the present invention does not need to move the pipette up and down or back and forth, thereby simplifying the structure of the device (see Fig. 35).

While the invention has been described using some preferred embodiments, these embodiments are illustrative and not restrictive. Those skilled in the art will appreciate that various changes and modifications can be made without departing from the spirit of the invention and the scope of the rights set forth in the appended claims.

The present invention provides a liquid flow device for performing a test including a biochemical reaction, a liquid metering device for automatically filling a liquid material into a liquid flow device, and various biological devices using the liquid flow device and the liquid metering device. Human diseases by providing a target substance extraction device for extracting target substances such as nucleic acids or proteins from a sample, and a target substance extraction method for extracting target substances from various biological samples using magnetic particles that selectively adsorb the target substances. It can be used in various fields such as the manipulation and biochemical analysis of biological samples, the development of new drugs, the examination of viral or bacterial infections, and the forensic science for the purpose of healing or preventing them.

Claims (26)

1. A plurality of storage chambers storing liquid materials;

   A reaction chamber for reacting two or more liquid materials;

   A plurality of feed transfer channels formed between the storage chamber and the reaction chamber for supplying the liquid material stored in the storage chamber to the reaction chamber;

   An injection channel formed to inject a liquid material into the plurality of storage chambers;

   And a discharge channel formed to be connected to the reaction chamber.

   By operating the pump means connected to the discharge channel and selectively opening and closing the plurality of injection channels to supply the liquid material stored in the storage chamber to the reaction chamber or to discharge the liquid material in the reaction chamber to the outside Liquid flow plate.
2. The method of claim 1,

   The liquid flow plate,

   A mixing chamber formed spaced apart from the reaction chamber;

   A mixed transfer channel configured to suck the liquid material in the reaction chamber into the mixing chamber or to discharge the liquid material in the mixing chamber into the reaction chamber;

   And an air suction channel formed at an upper end of the mixing chamber so as to suck air in the mixing chamber or inject air into the mixing chamber.

   By operating the pump means connected to the air suction channel and selectively opening and closing the plurality of injection channels to suck the liquid material stored in the storage chamber into the mixing chamber or to discharge the liquid material of the mixing chamber to the reaction chamber A liquid flow plate, characterized in that the material is mixed.
3. The method of claim 2,

   The liquid flow plate,

   An eluate chamber formed spaced apart from the reaction chamber;

   An eluate transfer channel formed to transfer the eluate in the reaction chamber to the eluate chamber;

   And an eluate recovery channel formed in the eluate chamber to suck the eluate.

   And operating the pump means connected to the eluate recovery channel and selectively opening and closing the plurality of injection channels to inhale or discharge the eluate stored in the storage chamber to the eluate chamber or to the outside.
4. The method of claim 1,

   The lower portion of the reaction chamber is a liquid flow plate further characterized in that the installation portion for allowing the magnetic application means or the heater close.
5. The method of claim 4, wherein

   The liquid flow plate,

   A second reaction chamber formed spaced apart from the reaction chamber and connected to the storage chamber through the feed transfer channel;

   A second eluate transfer channel formed to transfer the eluate in the reaction chamber to the second reaction chamber;

   And a liquid material suction channel formed to be connected to the second reaction chamber.

   Operating the pump means connected to the eluate recovery channel and selectively opening and closing the plurality of injection channels to suck the eluate from the reaction chamber and supply the liquid substance stored in the storage chamber to the second reaction chamber for reaction. Liquid flow plate.
6. The method of claim 5,

   The second reaction chamber may include: a second installation part allowing access of a heater or magnetic application means;

   And a viewing window for observing a reaction inside the second reaction chamber.
7. A plurality of storage chambers for storing a liquid material, an injection channel formed to inject liquid material into the storage chamber, a reaction chamber spaced apart from the storage chamber, and a liquid material stored in the plurality of storage chambers A plurality of feed transfer channels connected to supply the reaction chamber to the reaction chamber, a mixing chamber formed spaced at one side of the reaction chamber, and connected to the reaction chamber through a mixing transfer channel, and connected to a lower end of the mixing chamber to mix In the liquid flow block consisting of a plurality of unit liquid flow plate comprising a discharge channel for discharging the liquid material in the chamber and an air suction channel connected to the upper end of the mixing chamber for discharging the air in the mixing chamber,

   The injection channel is formed through the unit liquid flow plate;

   The plurality of unit liquid flow plate is a liquid flow block characterized in that the injection channel of the neighboring unit liquid flow plate is laminated so that the arrangement in the same line.
8. The method of claim 7, wherein

   The lower portion of the plurality of reaction chambers liquid flow block characterized in that it further comprises an installation portion for allowing the magnetic application means or the heater to be close.
9. The method of claim 8,

   The liquid flow block,

   A second reaction chamber formed spaced apart from the reaction chamber and connected to the storage chamber through the feed transfer channel;

   A second eluate transfer channel formed to transfer the eluate in the reaction chamber to the second reaction chamber;

   And a liquid material intake channel formed to be connected to the second reaction chamber.
10. The method of claim 8,

    The liquid flow block,

    An eluate chamber formed spaced apart from the reaction chamber;

    An eluate transfer channel formed to transfer the eluate in the reaction chamber to the eluate chamber;

    And an eluate recovery channel formed in the eluate chamber to suck the eluate.
11. In the target substance extraction apparatus using the liquid flow plate of any one of Claims 1-6,

    The target material extraction device is mounted to the liquid flow plate,

    Injection channel opening and closing means for selectively opening and closing a plurality of injection channels formed in the liquid flow plate;

    Pump means connected to the discharge channel, the air suction channel, and the eluate recovery channel formed in the liquid flow plate to suck the liquid substance or the eluate in the reaction chamber, the mixing chamber or the eluate chamber;

    Magnetic application means for fixing the magnetic particles to the inner wall of the reaction chamber;

    And a control means for controlling the injection channel opening and closing means, the pump means and the magnetic application means.
12. A target material extraction apparatus using the liquid flow block according to any one of claims 7 to 10,

    The target material extraction device is mounted to the liquid flow block,

    A main body having a mounting portion for mounting the liquid flow block;

    Liquid material and air supply means for injecting a liquid material through a plurality of injection channels formed on the side of the liquid flow block in close contact with the liquid flow block mounted on the mounting portion;

    Liquid material and air suction means for sucking liquid material and air through a plurality of injection channels formed on the other side of the liquid flow block;

    Liquid material conveying and discharging means for discharging the liquid material through the plurality of discharge channels and the air suction channels formed on the other side of the liquid flow block, or mixing the liquid material by sucking or pressurizing air;

    And an eluting liquid transporting and discharging means for transporting or discharging the eluent through a plurality of eluent recovery channels formed on another side of the liquid flow block.
13. The method of claim 12,

    The target material extraction apparatus includes magnetic application means for agglomerating and fixing magnetic materials on inner walls of the plurality of reaction chambers;

    And a control means for controlling the magnetic application means, liquid material and air supply means, liquid material and air suction means, liquid material transport and discharge means, and eluent transport and discharge means. Target material extraction device used.
14. The method of claim 12,

    The liquid material and air supply means,

    A plurality of main supply pipes tightly coupled to a plurality of injection channels formed in the liquid flow block;

    A plurality of liquid material tanks for storing the liquid material;

    A liquid material supply pipe connecting the main supply pipe and the liquid material tank;

    A liquid substance control valve installed in the plurality of liquid substance supply pipes;

    A plurality of air supply pipes connected to the plurality of main supply pipes and the other end of which communicates with the outside air;

    And a plurality of air supply control valves installed in the air supply pipe to regulate the flow of air.
15. The method of claim 14,

    The liquid material and air suction means,

    A plurality of main suction pipes tightly coupled to a plurality of injection channels formed in the liquid flow block;

    A plurality of main pumps installed in the main suction pipe to form a vacuum in the injection channel;

    A liquid material drain pipe connected to the main pump to discharge liquid material;

    And a plurality of main control valves installed on the plurality of main suction pipes to open and close the main suction pipes.
16. The method of claim 15,

    The liquid material transfer and discharge means,

    A plurality of liquid material discharge pipes connected to a plurality of discharge channels provided on another side of the liquid flow block;

    A main liquid material discharge pipe connected to the plurality of liquid material discharge pipes;

    A discharge pump installed at the main liquid material discharge pipe;

    A liquid substance discharge control valve installed in the main liquid substance discharge pipe;

    A liquid material drain pipe connected to the discharge pump;

    A plurality of air discharge pipes connected to the air suction and discharge channels of the liquid flow block;

    A main air discharge pipe connected to the plurality of air discharge pipes;

    A mixing pump installed at the main air discharge pipe;

    An air exhaust pipe connected to the mixing pump;

    Target material extraction apparatus using a liquid flow block comprising an air discharge control valve installed in the main air discharge pipe.
17. The method of claim 16,

    The eluate transfer and recovery means,

    A plurality of eluate discharge pipes connected to a plurality of air suction channels provided on another side of the liquid flow block;

    A main eluate discharge pipe connected to the plurality of eluate discharge pipes;

    An eluent pump connected to the main eluent discharge pipe;

    The target material extraction apparatus using a liquid flow block is installed in the main eluate discharge pipe and comprises an eluent discharge control valve for controlling the discharge of the eluate.
18. In the target material extraction method of extracting a nucleic acid material from a biological sample using the target material extraction apparatus equipped with the liquid flow plate according to claim 11,

    The target material extraction method,

    The liquid flow plate filled with the liquid material in the plurality of storage chambers is seated on the mounting unit provided in the target material extraction device, and the valve body of the injection channel opening and closing means provided in the target material extraction device is brought into close contact with a corresponding injection channel. A liquid flow plate mounting step of connecting means to the discharge channel, the air suction channel and the eluate recovery channel of the liquid flow plate;

    A liquid material supplying step of controlling the injection channel opening and closing means and the pump means to selectively supply the liquid material stored in the storage chamber of the liquid flow plate to the reaction chamber;

    Controlling the injection channel opening and closing means and the pump means to mix the liquid substance supplied to the reaction chamber or to disperse solid particles such as magnetic particles;

    A magnetic particle fixing step of agglomerating the dispersed magnetic particles inside the reaction chamber by using the magnetic applying means and attaching the magnetic particles to the inner wall of the reaction chamber;

    Controlling the injection channel opening and closing means and the pump means to discharge the remaining liquid in the reaction chamber to the outside;

    Controlling the injection channel opening and closing means and the pump means to transfer the eluate in the reaction chamber to the eluent chamber, or the eluent transfer and recovery step of recovering the eluent to the external storage container through the eluate recovery channel. Target material extraction method using a target material extraction device equipped with a flow plate.
19. A target material extraction method for extracting a nucleic acid material from a biological sample by using the target material extraction device equipped with the liquid flow block according to any one of claims 12 to 17,

    The target material extraction method,

    The liquid flow that seats the liquid flow block on the mounting portion provided in the main body and in close contact with the liquid material and air supply means, the liquid material and air suction means, the liquid material transfer and discharge means and the eluent transfer and discharge means provided around the mounting portion Block mounting step;

    A liquid material filling step of injecting a liquid material into each of the storage chambers through the plurality of injection channels provided in the liquid flow block by using the liquid material and air suction means and the liquid material and air supply means;

    A liquid material supplying step of selectively supplying the liquid material stored in the storage chamber of the liquid flow block to the reaction chamber by using the liquid material transport and discharge means and the liquid material and air supply means;

    By mixing the liquid material supplied to the reaction chamber by the liquid material transfer and discharge means and the liquid material and air supply means to the mixing chamber or by discharging the liquid material of the mixing chamber into the reaction chamber to mix two or more liquid materials A liquid material mixing step of dispersing solid particles such as magnetic particles;

    A magnetic particle fixing step of aggregating the dispersed magnetic particles inside the reaction chamber by using the magnetic application means and fixing the magnetic particles to the inner wall of the reaction chamber;

    A liquid material discharge step of discharging the remaining solution inside the reaction chamber and the mixing chamber to the outside by using the liquid material transfer and discharge means and the liquid material and air supply means;

    Eluent transfer and recovery step of transferring the eluate in the reaction chamber to the eluent chamber by using the eluent transfer and discharge means and the liquid material and air supply means or recovers to an external storage container through the eluent discharge channel Target material extraction method using a target material extraction device equipped with a liquid flow block.
20. The method of claim 19,

    The liquid material filling step,

    By operating a plurality of main pumps provided in the liquid material and the air suction means to form a vacuum in the main suction pipe, and close the plurality of air control valves provided in the liquid material and the air supply means, and connect with the liquid material storage tank. All the liquid material control valves are opened, and the liquid material of the liquid material storage tank is transferred through the injection channel by a vacuum formed inside the injection channel of the liquid flow block, and the liquid material to be transferred is injected into the injection channel. A liquid material injection step of sequentially introducing a plurality of storage chambers connected to the storage chambers and filling liquid materials into respective storage chambers and injection channels;

    By operating a plurality of main pumps provided in the liquid material and air suction means to form a vacuum in the main suction pipe, and at the same time, the plurality of liquid material control valves provided in the liquid material and air supply means are all closed and connected to the outside air. The air control valves of the plurality of air supply pipes are opened to allow external air to be sucked into the main suction pipe through the injection channel, until the vacuum formed in the injection channel is parallel to the surface tension and gravity inside the storage chamber. Using a target material extraction device equipped with a liquid flow block, characterized in that the liquid flow block is made of an air injection step of maintaining a constant level of the liquid material in each storage chamber by discharging the liquid material stored in the storage chamber to the outside through the injection channel Target material extraction method.
21. A plurality of injection channels penetrating from one side to the other side of the block;

    A plurality of storage chambers installed at predetermined intervals along the injection channel and having a neck portion connected to the injection channel at an upper end thereof;

    It is connected to the lower end of the plurality of storage chambers and the end is configured to include a plurality of discharge channels formed to be connected to the outside air,

    The liquid quantitative flow block of claim 1, wherein the liquid material stored in the storage chamber is discharged to the discharge channel by sucking the liquid material through the pump means connected to the injection channel or by sucking or discharging air.
22. A liquid quantitative supply device for quantitatively supplying a liquid substance using the liquid quantitative flow block according to claim 21,

    The metered supply device,

    Liquid material and air supply means installed on one side of the liquid metering flow block and connected to one side of the injection channel;

    And a liquid material and air suction means installed on the other side of the liquid metering flow block and connected to the other side of the injection channel.

    The liquid material and air supply means may include a plurality of main supply pipes tightly coupled to a plurality of injection channels formed in the liquid metering flow block, a plurality of liquid material tanks storing liquid materials, and the main supply pipe and liquid material tanks A liquid material supply pipe for supplying a liquid material by connecting a liquid material control valve installed in the plurality of liquid material supply pipes, a plurality of air supply pipes connected to the plurality of main supply pipes and communicating with an outside air, and the air A plurality of air supply control valves installed in the supply pipe to regulate the flow of air;

    The liquid material and air suction means, a plurality of main suction pipes are in close contact with the plurality of injection channels formed on the other side of the liquid metering flow block, and are installed to be connected to the main suction pipe to form a vacuum in the injection channel A plurality of main pumps, a liquid material drain pipe connected to the main pump to discharge liquid substances, a plurality of main control valves installed at the plurality of main suction pipes to open and close a main suction pipe, and connected to the main pump An apparatus for supplying a liquid, comprising an air discharge pipe for discharging air and an air discharge control valve installed in the air discharge pipe.
23. In the liquid quantitative supply method using the liquid quantitative supply device according to claim 22,

    The liquid quantitative supply method,

    Filling a liquid material into a plurality of injection channels formed in the liquid metering flow block and a plurality of storage chambers connected to the injection channels;

    An air suction step of sucking air in the injection channel so as to equalize the level of the liquid material injected into the plurality of storage chambers;

    And a liquid material discharge step of discharging the liquid material stored in the storage chamber to the outside.
24. The method of claim 23, wherein

    The liquid material filling step,

    By operating the main suction pump connected to the injection channel of the liquid metering flow block to form a low pressure inside the main suction passage and the injection channel, it is installed in the air supply pipe connected to the injection channel of the liquid metering flow block The liquid material control valve installed in the liquid material supply pipe connected to the injection channel is opened while the air control valve is closed, and the liquid material of the liquid material tank flows into the injection channel through the main supply pipe and flows into the injection channel. The injected liquid material is filled in a liquid supply method, characterized in that the storage chamber connected to the injection channel in turn.
25. The method of claim 24,

    The air suction step,

    By operating the main suction pump connected to the injection channel of the liquid metering flow block to form a low pressure in the main suction passage and the injection channel, the air control valve in a state in which the liquid material control valve is closed Opening the liquid, characterized in that the air is injected into the injection channel through the main supply pipe to be discharged to the outside through the main suction pipe and the main drain pipe.
26. The method of claim 25,

    In the liquid material discharge step, by operating the main suction pump connected to the injection channel of the liquid metering flow block to form a high pressure in the main suction passage and the injection channel, the liquid material control valve and air Closing the control valve, the high pressure is formed in the storage chamber and the liquid material stored in the storage chamber is discharged to the container or multi-well plate kit through the discharge channel, characterized in that the liquid supply.

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