WO2016025726A1

WO2016025726A1 - An analytic membrane array, and plasma separation device incorporating the same

Info

Publication number: WO2016025726A1
Application number: PCT/US2015/045077
Authority: WO
Inventors: Timothy Murray; Paul Henry NELSON; Daniel R. MCCLERNON; Anita Mcclernon
Original assignee: Vivebio, Llc
Priority date: 2014-08-13
Filing date: 2015-08-13
Publication date: 2016-02-18

Abstract

An analytic membrane array containing: a planar separator membrane having a porosity that gradually decreases from an upstream side to a downstream side of the membrane so as to filter and trap solid components of a biological material deposited on the separator membrane; and a planar capture membrane that allows for flow of a liquid component of the biological material therethrough. The capture membrane separably overlaps with at least a portion of the separator membrane at a downstream side of the separator membrane to provide for vertical or lateral downstream flow of the liquid component of the biological material from the separator membrane to the capture membrane for further filtering. A vertical flow separation device and a lateral flow separation device using the analytic membrane array for storage, preservation, and filtering of blood components, and separation of plasma are also provided.

Description

AN ANALYTIC MEMBRANE ARRAY, AND PLASMA SEPARATION DEVICE

INCORPORATING THE SAME

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application Nos. 62/036,985 filed August 13, 2014, 62/102,366 filed January 12, 2015, and 62/192,399 filed July 14, 2015, the entire contents of which are incorporated by reference herewith.

FIELD OF INVENTION

[0002] Embodiments generally relate to a biological specimen analytic membrane array, a device incorporating the analytic membrane array, and to methods for use therewith. More specifically, embodiments relate to an analytic membrane array for collection, separation, storage, and recovery of plasma from whole blood for subsequent quantitative and qualitative analysis.

BACKGROUND OF THE INVENTION

[0003] Biological specimens are often collected, transported and stored for analysis of the levels and concentrations of various analytes contained therewithin. Conventionally, liquid suspensions of biological specimens are stored in sealed airtight tubes under refrigeration. Liquid sample collection, handling, transportation and storage has many problems associated with it, for example: the cost of refrigeration (typically by dry ice) in remote collection centers; the risk of container breakage or leakage which causes loss of sample and the danger of infection; sample instability during shipment and storage; refusal of transport carriers to accept liquid biohazard shipments; and collection of adequate sample volume to ensure quantities compatible with laboratory methods of subsequent qualitative and quantitative analyses. The costs of addressing the above problems are substantial.

[0004] Dried blood spot (DBS) and dried plasma spot (DPS) sampling on filter paper are alternative methods to the liquid sampling procedures, and have been used worldwide with some success. Since the 1980s, manufacturers such as Schleicher and Schuell Corp., Bio-Rad, Boehringer Mannheim Corp., and Whatman, Inc., have been producing filter papers for DBS and DPS sampling. In using these commercially available biological sampling filter paper systems, a blood or plasma spot is placed in one or more designated areas of the filter paper, allowed to dry, and then mailed along with a test request form to the laboratory. Commonly used filter papers are known to those of ordinary skill in the art, such as WHATMAN 3 MM, GF/CM30, GF/QA30, S&S 903, GB002, GB003, or GB004. Several categories of blotting materials for blood specimen collection are available, e.g., S&S 903 cellulose (wood or cotton derived) filter paper and WHATMAN glass fiber filter paper. However, certain disadvantages have been associated with these commercially available filter papers. Specifically, certain of these commercially available and commonly used materials lack characteristics which provide precision values and accuracy that are preferred for carrying out certain qualitative and quantitative biological assays.

[0005] Genetic material can be extracted and isolated from prior art DBSs in sufficient quantities for use in genetic analysis. For instance, DBS has been used for the detection of prenatal human immunodeficiency virus (HIV) infection by the polymerase chain reaction (PCR) (Cassol, et al., J. Clin Microbiol. 30 (12): 3039-42, 1992). DPS and DBS have also been used with limited success for HIV RNA detection and quantification (Cassol, et al., J. Clin. Microbiol. 35: 2795-2801 , 1997; Fiscus, et al., J. Clin. Microbiol. 36: 258-60, 1998; O'Shea, et al., AIDS 13: 630-1, 1999; Biggar, et al., J. Infec. Dis. 180 1838-43, 1999; Brambilla, et al., J. Clin. Microbiol. 41(5): 1888-93, 2003); HIV DNA detection and quantification (Panteleefe, et al., J. Clin. Microbiol. 37: 350-3, 1999; Nyambi, et al., J. Clin. Microbiol. 32: 2858-60, 1994); and HIV antibody detection (Evengard, et al., AIDS 3: 591-5, 1989; Gwinn, et al., JAMA 265 : 1704-08, 1991). HCV RNA detection and genotyping are also reported using DBS (Solmone et al., J. Clin. Microbio. 40 (9): 3512-14, 2002). Although these studies provide a good correlation with titers using DPS or DBS as compared with conventional liquid plasma samples, a loss of viral titers may occur after room temperature storage (Cassol, et al., J. Clin. Microbiol. 35: 2795-2801, 1997; Fiscus, et al., J. Clin. Microbiol. 36: 258-60, 1998). DBS and DPS samples are clearly less expensive and less hazardous to transport than liquid samples.

[0006] However, the procedure of analyte microextraction from DBS and DPS on filter paper suffers from a number of disadvantages. For example, microextraction of sufficient DNA or RNA from filter paper involves reconstitution in a liquid medium under certain vigorous procedures, e.g., vortex and centrifugation that damages the genetic analytes of interest. Furthermore, the fibers and other components of the filters become dislodged into the reconstitution solution, and require further centrifugation separation and/or can impede the ability to isolate the genetic material, such as by blocking genetic material from adhering to a separation column. Such prior microextraction procedures require a high standard of technical assistance, and even then do not consistently provide results with a desired level of sensitivity, reproducibility, quantification and specificity.

[0007] Furthermore, the sample volume used for DBS and DPS on filter paper is limited, typically to 50-200 μΐ spots, and imposes considerable difficulty in analyte detection. As a result, accurate quantification and reproducibility can be encountered, particularly when the concentration of the desired analyte material is low in the sample. Also in the prior art, there is a lack of deliberate inhibition of enzymes and chemicals which degrade the analytes, such as genetic material contained there within. Even in the presence of a bacteriostatic agent there are conditions that permit enzymatic, non- enzymatic and autolytic breakdown of the genetic material. Furthermore, microextraction of genetic material from DBS or DPS on filter papers is considerably more difficult if absorption of high molecular weight DNA or RNA is required. Although the introduction of new material and transportation methods continuously improve the ways samples are handled, the quantity and quality of the sample available for subsequent analysis are still of great concern to researchers and clinicians alike.

[0008] For example, U.S. Patent No. 7,638,099, incorporated by reference herein in its entirety, provides an advantageous alternative system for biological specimen collection, storage and transportation. The reference suggests the use of cellulose acetate fibers and hydrophilic polymer fibers as being advantageous for an absorbent matrix material. The use of whole blood creates technical challenges with certain matrix and the system because solid components of the whole blood, such as RBCs, WBCs, and other cellular components may clog the matrix, resulting in severe diminished recovery of the absorbed materials, such as plasma and/or virus comprising analytes of interests for subsequent analysis, e.g., viral load testing. Further improvements are desired for certain situations, such as to achieve more accurate and reproducible quantification of viral load in a sample.

[0009] There is a need for an improved device for collection, storage and transportation of liquid suspension of biological specimens, such as whole blood, containing analytes of interest in a dry state, especially in large field studies and for application in settings where collection, centrifugation, storage and shipment can be difficult, as is often the case in developing countries. There is also a need for an improved device with a membrane assembly that can filter out and/or trap solid components (e.g., white blood cells, red blood cells, etc.) of whole blood, as well as separate serum/plasma from the whole blood so as to improve the recovery of virus, plasma proteins, cytokines, chemokines, immunoglobins, etc. for subsequent analysis that provides precision values and accuracy of detection, reproducibility and quantification of the analytes of interest contained therewithin, and eliminate the need for centrifugation.

SUMMARY OF THE INVENTION

[0010] An analytic membrane array and a device containing the same according to embodiments described herein provide for a safe, convenient, and simple method for collecting, filtering, preserving, storing, and transporting biological specimens containing analytes of interest. An inventive use of the analytic membrane assembly further fulfills the need to recover biological specimens containing analytes of interest for subsequent analysis with improved sensitivity and specificity of detection.

[0011] In embodiments, an analytic membrane array comprises a planar asymmetric separator membrane and a planar capture membrane that separably overlaps with at least a portion of the separator membrane at a downstream side of the separator membrane so as to provide for vertical or lateral downstream flow of the liquid component of the biological sample (specimen) from the separator membrane to the capture membrane.

[0012] In certain embodiments, the planar capture membrane may be made of a material that allows for flow of a liquid component of the biological specimen therethrough, and the separator membrane may be made of polysulfone polymer material having a porosity that gradually decreases from an upstream side to a downstream side of the membrane so as to filter and trap solid components of a biological specimen deposited on the separator membrane.

[0013] The analytic membrane array may be used to trap solid components of a biological specimen, examples of the biological specimen including (but not limited to) whole blood, plasma, urine, saliva, sputum, semen, vaginal lavage, bone marrow and cerebrospinal fluid. In embodiments, the separator membrane is configured to filter and trap solid components of, e.g., a whole blood specimen, and the capture membrane is configured to separately filter and trap a plasma fraction or filtrate of the whole blood specimen.

[0014] The analytic membrane array according to embodiments of the invention may be a multi-spot membrane, configured to receive and store a plurality of biological specimens (i.e., "spots").

[0015] Embodiments also relate to a separator device comprising the analytic assembly described herein and the following:

(a) a support assembly of an elongated bifacial sheet having a first configuration in which the bifacial sheet is in a foldable state and a second configuration in which the bifacial sheet is in a folded state, wherein the bifacial sheet has an upper surface and an inner (i.e., lower) surface and is substantially equally divided into 4 foldable quadrants by 3 parallel crease lines in the bifacial sheet, the foldable quadrants including:

a first central quadrant and a second central quadrant mutually adjacent to a central crease line in the bifacial sheet, wherein the second central quadrant contains an aperture in the bifacial sheet to allow for deposition of a blood specimen therethrough, a first outer quadrant adjacent to the first central quadrant, and a second outer quadrant adjacent to the second central quadrant;

(b) the analytic membrane assembly disposed on the bifacial sheet, such that the separator membrane is adhered to the upper surface of the second central quadrant and covers the aperture therein and the capture membrane is adhered to the upper surface of the first central quadrant; and

(c) an adhesive strip affixed to the upper surface of the bifacial sheet along an outer perimeter of each of the first central quadrant and the second central quadrant, wherein the first outer quadrant is foldable onto the first central quadrant so as to place and optionally seal the upper surface of the first outer quadrant directly over the upper surface of the first central quadrant to enclose and protect the capture membrane located on the first central quadrant, and the second outer quadrant is foldable onto the second central quadrant so as to place and optionally seal the upper surface of the second outer quadrant directly over the upper surface of the second central quadrant to enclose and protect the separator membrane comprised on the second central quadrant.

[0016] In certain embodiments, the separator device is a vertical flow plasma separation device and may contain therein a removable desiccant paper near the separator membrane and/or the support membrane. In additional embodiments, the vertical flow plasma separation device may have a bifacial sheet having a folded state according to the second configuration of the support assembly that includes the first outer quadrant folded onto the first central quadrant so that the upper surface of the first outer quadrant is placed over the upper surface of the first central quadrant and sealed in place by the adhesive strip affixed to the upper surface of the bifacial sheet along the outer perimeter of the first central quadrant; and the second outer quadrant folded onto the second central quadrant so that the upper surface of the second outer quadrant is placed over the upper surface of the second central quadrant and sealed in place by the adhesive strip affixed to the upper surface of the bifacial sheet along the outer perimeter of the second central quadrant.

[0017] In certain embodiments, the vertical flow plasma separation device may contain the bifacial sheet having the folded state (according to the second configuration of the support assembly) folded onto the second central quadrant along the central crease line so as to provide the blood separation device in a closed state, the lower surface of the first outer quadrant that is already folded over the first central quadrant being placed over the lower surface of the second outer quadrant that is already folded over the second central quadrant, whereby the lower surface of each of the first and second central quadrants constitutes an outer surface of the vertical flow plasma separation device in a closed state.

[0018] In certain embodiments, the separator device is a lateral flow blood separation device. The lateral flow plasma separation device may contain:

(a) a support assembly of two sheets separably coupled together in a foldable configuration and in a folded configuration, the sheets including an elongated bifacial base sheet having an upper surface and an inner (i.e., lower) surface, and substantially equally divided into a first and second foldable panel by a central folding line, the first foldable panel being further divided into an outer quadrant and an inner quadrant that is between the outer quadrant of the first panel and the second panel, and an elongated bifacial cover sheet having an upper surface and a lower surface, a size corresponding to that of the first panel of the base sheet, and an aperture to allow for deposition of a blood specimen therethrough,

(b) the analytic membrane assembly disposed between the first panel of the base sheet and the cover sheet such that the separator membrane is located atop the outer quadrant of the first panel, wherein the separator membrane has a teardrop shape with an elongated portion near the downstream side thereof, the elongated portion of the separator membrane separably overlapping with at least a portion of the capture membrane;

(c) a layer of non-absorbent material adhered to a lower surface of the separator membrane; and

(d) an adhesive strip affixed to the upper surface of the base sheet along an outer perimeter of the first panel so as to optionally adhere the cover sheet to the first panel of the base sheet.

[0019] The lateral flow plasma separation device is furthermore separable from the capture membrane by adhesion to the lower surface of the cover sheet, whereby the separator membrane becomes adhered to the lower surface of the cover sheet placed thereon and separated from the capture membrane as a result of the cover sheet folding onto the second panel of the base sheet; and the outer quadrant is foldable onto the central quadrant so as to place and optionally seal the upper surface of the outer quadrant directly over the upper surface of the central quadrant to enclose and protect the capture membrane comprised on the central quadrant.

[0020] Certain embodiments also relate to a blood/plasma separation device that may comprise: a separator membrane having a porosity that gradually decreases from an upstream side to a downstream side so as to filter and trap solid components of a blood sample deposited on the separator membrane; and a plasma collection chamber separably adhered to a lower surface of the downstream side of the separation membrane, wherein the plasma collection chamber is configured to collect and store plasma that has been separated from a blood sample by flow from an upstream side to a downstream side of the separator membrane. In such embodiments, the plasma collection chamber may overlap with at least a portion of the separator membrane at a downstream side of the separator membrane so as to provide for vertical downstream flow of the liquid plasma component of the whole blood sample from the separator membrane to the plasma collection chamber for storage and/or further filtering.

[0021] In some certain embodiments, the blood/plasma separation device may comprise a multi-spot membrane array according to embodiments of the invention, so as to provide sample storage configurations on the card including sample spot arrangements of, e.g., 1 x 2 (one row of two sample spots), 2 x 2 (two rows of two sample spots), 2 x 3 (two rows of three sample spots), 2 x 4 (two rows of four sample spots), configuration, or 1 x 4 (one row of 4 sample spots).

[0022] In embodiments, a blood/plasma separation device according to the invention may comprise:

(a) a support assembly of an elongated bifacial sheet having a first configuration in which the bifacial sheet is in a foldable state and a second configuration in which the bifacial sheet is in a folded state, wherein the bifacial sheet has an upper surface and an inner surface, and is substantially equally divided into two foldable quadrants by a central parallel crease line in the bifacial sheet, the foldable quadrants including a first quadrant and a second central quadrant that are mutually adjacent to the central crease line in the bifacial sheet;

(b) a plurality of spots provided on the first quadrant for deposition and storage of whole blood specimens of a patient;

(c) a plurality of spots provided on the second quadrant for deposition and storage of plasma specimens of the patient; and

(d) an adhesive strip affixed to the upper surface of the bifacial sheet along an outer perimeter of the second quadrant, wherein the first outer quadrant is foldable onto the first central quadrant so as to place and optionally seal the upper surface of the first quadrant directly over the upper surface of the second quadrant to enclose and protect the specimens deposited therein.

[0023] In certain embodiments, the blood/plasma separator device may include a tab extending from opposite end sides of each of the first and second quadrants so as to enable ease of folding together and subsequent separation of the first and second quadrants from each other. In some embodiments, the plurality of spots provided on the first quadrant contain raised circular perimeters to facilitate a determination whether an appropriate volume of whole blood has been deposited thereon.

[0024] In embodiments, the first and second quadrants of the blood plasma separator device may each include from 2-4 spots arranged in one or more rows. In further embodiments, the length of the first and second quadrants constituting the elongated bifacial sheet according to the first configuration may be from about 50 mm to about 250 mm, and a width of the elongated bifacial sheet constituted by the quadrants may be from about 20 mm to about 60 mm.

[0025] In certain embodiments of the invention, the blood/plasma separator device may also include a separate and independently foldable quadrant for deposition and storage of additional whole blood or plasma specimens for further analysis.

[0026] The blood/plasma separation device according to the invention may alternatively comprise a plasma card separate from the whole blood deposited on the separator membrane.

[0027] The blood/plasma separation device according to the invention may alternatively comprise a plasma card separate from the whole blood deposited on the separator membrane. In some certain embodiments, the blood/plasma separation device may comprise a multi-spot membrane array according to embodiments of the invention, so as to provide sample storage configurations on the card including sample spot arrangements of, e.g., 1 x 2 (one row of two sample spots), 2 x 2 (two rows of two sample spots), 2 x 3 (two rows of three sample spots), 2 x 4 (two rows of four sample spots), configuration, or 1 x 4 (one row of 4 sample spots).

[0028] The blood/plasma separation device is preferably a point-of-care device and easily portable. As described further herein, the blood/plasma separation device may further include an identification portion on outer surface thereof when in a fully folded/closed configuration for identifying the associated patient with the biological samples collected and stored therein. The identification portion may take the form of a label that identifies the patient, and/or secure barcodes or images. The identification portion may furthermore be configured to allow for scanning, transmission, and cloud- based storage of the identifying information and/or subsequent authentication by a smartphone application. In embodiments, the identification portion includes identification technology that provides for HIPAA-compliant cloud-based transmission and storage of personal identifying information of a patient associated with the biological specimens stored in the device. In certain embodiments, the identification technology comprises an authentication feature that enables authentication of specimens stored in the device.

BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The manner in which objectives of the present disclosure and other desirable characteristics may be obtained will become further evident from the following descriptions of the appended drawings.

[0030] FIGS. 1 A-1C show a polysulfone asymmetric polymer used as a separation membrane in the analytic membrane array for blood component filtering and serum/plasma separation from whole blood according to embodiments of the invention. FIG. IB shows captured red cells, whereas FIG. 1C shows the separated cell- free plasma. The arrow in FIG. 1A shows the direction of decreasing pore size from a large pore region to a fine pore region of the separation membrane.

[0031] FIG. 2 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in an unfolded (foldable) configuration.

[0032] FIG. 3A is a cross-sectional view and FIG. 3B is a perspective view of an embodiment of a vertical-flow blood/plasma separator device of the invention in a foldable, but already partly folded, configuration.

[0033] FIG. 4 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0034] FIG. 5 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0035] FIG. 6 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0036] FIG. 7 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration. [0037] FIG. 8 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0038] FIG. 9 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0039] FIG. 10 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0040] FIG. 11 is a perspective view of an embodiment of a vertical- flow blood/plasma separator device in a folded configuration.

[0041] FIG. 12 is a perspective view of an embodiment of a vertical-flow blood/plasma separator device in an unfolded configuration as shown in Fig. 2, but also including desiccant paper in close proximity to the analytic membrane array.

[0042] FIG. 13 is a perspective view of an embodiment of the analytic membrane array assembly within a lateral-flow blood/plasma separator device in an unfolded (foldable) configuration.

[0043] FIG. 14 is a perspective view of an embodiment of the analytic membrane array assembly within a lateral-flow blood/plasma separator device.

[0044] FIG. 15 is a perspective view of an embodiment of a lateral- flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0045] FIG. 16 is a perspective view of an embodiment of a lateral- flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0046] FIG. 17 is a perspective view of an embodiment of a lateral- flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0047] FIG. 18 is a perspective view of an embodiment of a lateral- flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0048] FIG. 19 is a perspective view of an embodiment of a lateral- flow blood/plasma separator device in a foldable, but already partly folded, configuration.

[0049] FIG. 20 is a perspective view of an embodiment of a lateral-flow blood/plasma separator device in an almost-folded configuration. [0050] FIG. 21 is a perspective view of an embodiment of a lateral- flow blood/plasma separator device in an almost-folded configuration.

[0051] FIG. 22 is a perspective view of an embodiment of a lateral-flow blood/plasma separator device in an unfolded (foldable) configuration as shown in FIG. 13, but including desiccant paper.

[0052] FIG. 23 is a perspective view of an embodiment of a lateral-flow blood/plasma separator device in a folded configuration.

[0053] FIG. 24 is a perspective view of a multiple spot blood/plasma separator device in an unfolded (foldable) configuration according to another embodiment of the invention.

[0054] FIG. 25 is a perspective view of a multiple spot blood/plasma separator device in an unfolded (foldable) configuration according to another embodiment of the invention.

[0055] FIG. 26 is a perspective view of a multiple spot blood/plasma separator device in an unfolded (foldable) configuration according to another embodiment of the invention.

[0056] FIG. 27 is a perspective view of a multiple spot blood/plasma separator device in a partly folded configuration according to another embodiment of the invention.

[0057] FIG. 28 is a perspective view of a multiple spot blood/plasma separator device in a partly folded configuration according to another embodiment of the invention.

[0058] FIGS. 29A-29C show perspective views of a blood/plasma separator device with an identification/security feature on an outside surface thereof according to various embodiments of the invention. Specifically, FIG. 29A shows the feature as RFID, FIG. 29B shows the feature as a 2D barcode, and FIG. 29C shows a TRAXSECUR information and barcoded label for use in connection with the exemplified smartphone application.

[0059] FIGS. 30A-30C show smartphone interfaces of the exemplified

TRAXSECUR application for authentication of biological specimens obtained using the device of the invention. DETAILED DESCRIPTION OF THE INVENTION

[0060] Throughout this application, various publications are referenced. The disclosures of all referenced publications, as well as those referenced within those publications, are hereby incorporated by reference herein in order to more fully describe the state of the art to which the present disclosure pertains.

[0061] As used herein, the terms "a" or "an" mean one or more than one depending upon the context in which they are used. For example, "an analyte" in a sample refers to a particular type of analyte of interest (such as, e.g., intact HCV or HIV RNA), of which there may be numerous copies within the sample. Where a sample is referred to as containing an analyte, it is understood that the sample may contain many other types of analytes of interest also. Throughout the specification, the terms "comprise," "comprises," "comprising" and the like shall consistently mean a collection of applicable features, and should not be limited to those objects.

[0062] As used herein, the term "analytes of interest" refers to any micro- or macro- molecules in the biological specimen that are interested to be detected or analyzed. These include, for example, nucleic acids, polynucleotides, oligonucleotides, proteins, polypeptides, oligopeptides, enzymes, amino acids, receptors, carbohydrates, lipids, cells, any intra- or extra- cellular molecules and fragments, virus, viral molecules and fragments, or the like. In certain embodiments, the analytes of interest can be exogenous natural or synthetic compounds, such as small molecules, like drugs, prodrugs or metabolites thereof. In certain embodiments, the analytes of interest are nucleic acids including either or both proviral and/or viral DNA or RNA. As used herein, the term "nucleic acids" or "polynucleotide" refers to RNA or DNA that is linear or branched, single or double stranded, a hybrid, or a fragment thereof. The term also encompasses RNA/DNA hybrids. The term also encompasses coding regions as well as upstream or downstream noncoding regions. In addition, polynucleotides containing less common bases, such as inosine, 5- methylcytosine, 6-methyladenine, hypoxanthine, and other are also encompassed. Other modifications, such as modification to the phosphodiester backbone, or the 2'-hydroxy in the ribose sugar group of the RNA are also included. The nucleic acids/polynucleotides may be produced by any means, including genomic preparations, cDNA preparations, in vitro synthesis, RT-PCR, and in vitro or in vivo transcription. In certain embodiment, the nucleic acids are either or both proviral and/or viral DNA or RNA, for example, proviral or viral DNA or RNA from human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), or any other human or animal viral pathogen. In certain embodiments, the analytes of interest are viral particles for determining viral load, and biological markers for determining HLA blood types, useful for molecular diagnostic genotyping.

[0063] "Continuous" means that the relevant components of the claimed invention are connected at all times during use. For exemplary purposes only, the drawings show a unitary base of the low-profile or flat, planar type, whose height is significantly smaller than its width and depth. This base may be used to house or store a biological specimen as described herein. However, the principles described herein are equally applicable to other types of "continuous" sheets including those where the height is comparable to or greater than the width or depth of the base sheet.

[0064] The term "absorb" and "adsorb" are used interchangeably, and means that the liquid suspension is incorporated into or onto the two-portion matrix in such a way as to be readily removed from the matrix while leaving the analytes of interest behind.

[0065] As used herein, the term "biological specimen" refers to samples, either in liquid or solid form, having dissolved, suspended, mixed or otherwise contained therein, any analytes of interest, such as, e.g., genetic material. As used herein, the term "genetic material" refers to nucleic acids that include either or both deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). The term "biological specimen" also refers to whole blood, plasma, serum, lymph, synovial fluid, bone marrow, cerebrospinal cord fluid, semen, saliva, urine, feces, sputum, vaginal lavage, skin scrapings, hair root cells, or the like of humans or animals, physiological and pathological body liquids, such as secretions, excretions, exudates and transudates; any cells or cell components of humans, animals, plants, bacteria, fungi, plasmids, viruses, parasites, or the like that contain analytes of interest, and any combination thereof.

[0066] As used herein, the term "compress," "compressible," "compression," and other derivatives of the word "compress" means that the volume of the saturated analytical membrane array reduced as compared to the original volume of the saturated assembly while force or a pressure is applied thereto. As used herein, the term "a portion of the biological specimen" means at least some of the biological specimen contained in the liquid suspension is released from each portion of the matrix. In certain embodiments, the analytical membrane array of the invention can be physically separated from each other and each portion of the membrane array having the captured components thereon can be reconstituted with a reconstitution media and then be separately compressed until the maximum volume of the reconstituted components from the biological specimen is released from the matrix.

[0067] As used herein, the term "liquid suspension" refers to any liquid medium and mixture containing biological specimens. This includes, for example, water, saline; cell suspensions of humans, animals and plants; extracts or suspensions of bacteria, fungi, plasmids, viruses; extracts or suspensions of parasites including helminthes, protozoas, spirochetes; liquid extracts or homogenates of human or animal body tissues, e.g., bone, liver, kidney, brain; media from DNA or RNA synthesis; mixtures of chemically or biochemically synthesized DNA or RNA, and any other sources in which any biological specimen is or can be in a liquid medium.

[0068] As used herein, the term "oxygen scavenging element" refers to is a substance that consumes, depletes or reduces the amount of oxygen from a given environment without negatively affecting the samples of interests.

[0069] For ease of description, separation devices and various components used therewith are described herein in their usual assembled positions (folded and unfolded) as shown in the accompanying figures. Terms such as "upper," "lower," "vertical," "longitudinal," etc., are used herein with reference to these usual positions. However, the separation devices and the analytic membrane arrays may be manufactured, transported, sold or used in orientations other than those described and shown herein.

[0070] The invention may be understood more readily by reference to the following detailed description of various embodiments of the invention and the examples included herein. However, before the present devices, materials, and methods are disclosed and described, it is to be understood that this invention is not limited to specific embodiments of the devices, materials and methods, as such may, of course, vary, and the numerous modifications and variations therein will be apparent to those skilled in the art. It is also to be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting. [0071] Embodiments described herein generally relate to an analytic membrane array and a device, such as a blood separator, containing the analytic membrane array, as well as to methods of use of the analytic membrane array and a separator device containing the same for collection, filtration, storage, and transportation of a liquid suspension of a biological specimen containing an analyte of interest. Additionally, certain embodiment relate to an analytic membrane array as described herein, and to methods of use, collection, filtration, storage, and transportation of a liquid suspension containing a biological specimen in a dry state that is convenient and simple to use.

[0072] More particularly, embodiments of the invention relate to a dual layer analytic membrane array. Although the analytic membrane array is suited for use in a separation device as described in embodiments herein, other uses will become evident to those of ordinary skill in the art. One role of the analytic membrane array is to separate red blood cells or erythrocytes from whole blood in a separation membrane. Another role of the analytic membrane array is to have enough capillary action so that separated serum or plasma flows from the separation membrane to the capture membrane. In yet another device according to embodiments of the invention, an analytic membrane array provides for separate storage of whole blood and plasma specimens. In alternative embodiments, also provided is an analytic membrane array for direct application of plasma and whole blood samples, respectively, in separate quadrants thereof.

[0073] An analytic membrane array and a device containing the same according to embodiments of the invention provide for a safe, convenient, and simple method for collecting, filtering, preserving, storing, and/or transporting biological specimens containing analytes of interest. The convenient and fully integrated "matchbox" configuration of the blood/plasma separation device provided in certain embodiments of the invention is suitable for use as a point-of-care device for collection and storage of biological specimens containing analytes of interest from a patient. A further advantage of the disclosed analytic membrane array for use in a separation device is a decrease in hemolysis of red blood cells as compared to other blood separation media. Excessive hemolysis can make the plasma appear red instead of clear/yellowish in color.

[0074] Variables that tend to influence the performance of blood/plasma separation media include, for example, the distance traveled by the whole blood and resulting serum, the time required for the blood to absorb, and protein binding on the surface of such media. Therefore, one of the goals of the present analytic membrane array is to provide a separation media comprising an upstream layer (the "separation membrane") and a downstream layer ("capture membrane") that overlap with each other, wherein the separation membrane is designed to separate the white and red blood cells from the whole blood and the capture membrane is designed to allow for a fluid portion of the whole blood sample to flow therethrough. Specifically, the membrane array according to the present disclosure comprises a separation membrane for filtering and/or trapping cellular material, removably attached to a capture membrane for filtering and/or trapping nucleic acids in plasma. In certain embodiments, the separation membrane is preferably an asymmetric porous membrane and the capture membrane is a cellulose fiber filter paper.

[0075] Referring to FIGS. 1A-1C, illustrating that the separation membrane has pore size that gradually decreases from an upstream side of the membrane to a downstream side in the direction of the arrow. When the whole blood flows through, the large molecule solid components (2), such as WBCs, RBCs, etc. in the whole blood, would be captured in the upstream side where it contains large pore size polymer, as shown in FIG. IB (captured red blood cells). The fluid components (4) containing cell- free serum, plasma, and plasma proteins can then be drawn through the membrane (in the direction of the arrow in FIG. 1A) via gravity and/or capillary action to the downstream side where it contains the finer smaller pore sized polymer, as shown in FIG. 1C (separated cell-free plasma). The asymmetric polysulfone polymer membrane portion of the inventive matrix thus provides blood components filtering and cell-free serum or plasma separation.

[0076] In embodiments, an analytic membrane array comprises a planar asymmetric separator membrane and a planar capture membrane that separably overlaps with at least a portion of the separator membrane at a downstream side of the separator membrane so as to provide for vertical or lateral downstream flow of the liquid component of the biological sample (specimen) from the separator membrane to the capture membrane.

[0077] The analytic membrane array may be used to trap solid components of a biological specimen, examples of the biological specimen including (but not limited to) whole blood, plasma, urine, saliva, sputum, semen, vaginal lavage, bone marrow and cerebrospinal fluid. In embodiments, the separator membrane is configured to filter and trap solid components of, for example, a whole blood specimen, and the capture membrane is configured to separately filter and trap a plasma fraction, filtrate, or plasma proteins of the whole blood specimen.

[0078] The planar capture membrane according to embodiments of the invention may be made of a material that allows for flow of a liquid component of the biological specimen therethrough and may comprise a plurality of fibers, and the separation membrane may be made of a material that has gradually decreasing pore size from an upstream side to a downstream side. In certain embodiments, flow of the liquid component of the biological specimen deposited on the separator membrane is effectuated by capillary flow.

[0079] Materials suitable for use in the separation membrane (an asymmetric porous membrane) are those through which plasma can move faster than corpuscles, such as, e.g., synthetic polymers having fine fiber diameter, fibers made of glass or porous polymers. A material of the separation membrane according to certain preferred embodiments may be made of polysulfone polymer material having a porosity that gradually decreases from an upstream side to a downstream side of the membrane so as to filter and trap solid components of a biological specimen deposited on the separator membrane. Exemplary separation membrane materials also include (but are not limited to) synthetic or natural polymers, such as cellulose mixed esters, polyvinylidene difluoride, polytetrafluoroethylene, polycarbonate, polypropylene, polyester, polysulfone polymers and matrices (e.g., Asymmetric Sub-Micron Polysulfone (BTS) and/or Asymmetric Super Micron Polysulfone (MMM) made by Pall Corporation). In certain embodiments, the material of the separator membrane may be VIVID GR, VIVID GX, and CYTOSTEPH 1660. However, any asymmetric membrane or filtering material now known and/or commercially available or later developed, that is suitable for blood component filtering and serum/plasma separation is contemplated as being within the scope of the disclosure. In embodiments, the separation membrane has a porosity of not more than 30%, preferably not more than 25%. In certain embodiments, the separation membrane may be made of polysulfone polymer having a pore size ranging from, e.g., about 0.1-20 microns and a pore size ratio from about 50:1 to 100:1.

[0080] Materials suitable for use as the capture membrane according to embodiments of the invention may include, but are not limited to, polymers, cotton, and/or cellulose. Filter papers that may be selected for use as the capture membrane include cellulose fiber papers manufactured from cotton linters. Cotton linters (i.e., cotton wool) are short fibers that adhere to seeds of a cotton plant after the longer fibers have been pulled from the cotton seed. An exemplary material of the capture member may include AHLSTROM 222, which is a 100% cotton fiber. In certain embodiments, a majority of the cellulose fibers of a cellulose fiber filter paper used as the capture membrane may have sizes in the range of about 1 -100 microns, 10-50 microns, or 20-25 microns in length and may contain numerous hydrophobic pockets.

[0081] Exemplary filter papers suitable for use in the analytical membrane array according to certain embodiments include (but are not limited to) filter papers for blood collection registered by the U.S. Food and Drug Administration as Class II Medical Devices (21 CFR §862.1675). In embodiments, the capture membrane (i.e., the second layer of the analytical assembly array) may be the FDA-cleared/approved filter paper WHATMAN 903, AHLSTROM 142, AHLSTROM 226, AHLSTROM 222, Ahlstrom 270, and ESSENTRA.

[0082] In certain embodiments, each of the separation membrane and the capture membrane may have a shape, such as, e.g., a circle, oval, square, rectangle, triangle, or other shapes and surface textures suitable for absorption and use in assemblies described further herein. In certain embodiments, each membrane of the analytical assembly array may have the same or similar shape and/or the same or similar dimensions. Alternatively, the asymmetric porous membrane of the first portion may be different in size and/or shape than the cellulose fiber filter paper of the second portion.

[0083] In certain embodiments, a size of the separation membrane may be larger than a size of the separation membrane to which it is removably attached. For example, the size of the asymmetric porous membrane may be at least 20%, or at least 30%, or at least 40%, or at least 50% larger than a size of the cellulose fiber filter paper. In embodiments, the separation membrane may also have a shape that is different from a shape of the capture membrane and, thus, does not align in its entirety with the shape of the cellulose fiber paper if brought into contact for attachment thereto. For example, the separation membrane may have a shape that is substantially circular and larger in size than a size of the filter paper having, e.g., a circular shape. As a further example, the separation membrane may have an irregular or oblong shape (e.g., a racquet shape with a handle-like extension extending on a lateral side thereof), while the hydrophilic filter paper has a circular shape.

[0084] In certain embodiments, the separation membrane and the capture membrane may have diameters of from about 1mm to 50 mm, or from 10 mm to 30 mm, inclusive. For example, the separation membrane may have a diameter of 14 mm, 16 mm, 18 mm or 20 mm, and the capture membrane may have a diameter of 7 mm or 14 mm.

[0085] The analytical membrane array may be formed by, e.g., removably attaching the capture membrane (e.g., cellulose fiber filter paper) to the separation membrane (an asymmetric porous membrane) at any overlapping or contacting portions thereof. For example, the capture membrane may be in contact with and joined to the analytical membrane over an entire surface area thereof. Alternatively, the capture membrane may be in contact with a surface area of the separation membrane that is smaller than (and does not cover) the full surface area of cellulose fiber filter paper, as shown in FIG. 12. The separation membrane and the capture membrane of the analytical membrane array are configured to be separable from one another, and therefore can be stacked together at overlapping regions thereof with or without an adhesive or a surrounding sheath of material that may be perforated at their junction or adapted for being cut into two separate stages of a pre-defined indicia.

[0086] Embodiments also relate to a blood/plasma separation device that contains the analytic membrane array described herein. Alternatively, a blood/plasma separation device may contain the separator membrane (of the analytic membrane array described herein) positioned over a plasma collection chamber or vessel. In such embodiments, the plasma collection chamber or vessel (rather than the capture membrane of the analytic membrane array described herein) may be positioned so as to separably overlap with at least a portion of the separator membrane. The separation device may be a vertical flow separation device or a lateral flow separation device, as described in the exemplified embodiments below.

[0087] The analytical membrane array according may be removably integrated into a device, such as, e.g., a point-of-care device. That is, the blood/plasma separation device is able to analyze plasma that has been isolated from a whole blood sample retrieved from a patient at a point-of-care. The point-of-care blood/plasma separation device may utilize at least some aspects of the membrane array described herein to produce a plasma sample from a small quantity of whole blood. This can be done, e.g., in a doctor's office or at a patient's bedside without a power supply. The plasma sample collected and stored in the plasma collection chamber may be of a quality and composition that are comparable to plasma retrieved by centrifuging. Blood/plasma separation devices according to the present disclosure may be configured to separate components of a whole blood sample by means of a flow channel through one or more operations selected from centrifugation and/or capillarity flow.

[0088] Therefore, methods are also provided for collecting a volume of plasma from a liquid biological specimen obtained from a patient by utilizing the analytic membrane of the invention. Certain embodiments also relate to a system that includes means for collecting a volume of plasma. Such a system may include means for introducing a whole blood sample onto a separation membrane that filters and/or traps cellular material of the whole blood, and a means for extracting the plasma from the separation membrane into the plasma collection chamber, (e.g., by vertical downstream flow).

[0089] A plasma collection chamber or vessel as described herein may be of a shape, size, and material suitable for use in the collection and storage of a liquid plasma composition that could be determined by persons skilled in the art. The device may be, for example, a blood or plasma separator that allows for separate analysis of the blood cells and the plasma, respectively. The device comprising the analytical membrane array according to embodiments described herein may be a "vertical flow" separator or, alternatively, a "lateral flow" separator. Embodiments also relate to a separator device comprising the analytic assembly described herein and the following:

(a) a support assembly of an elongated bifacial sheet having a first configuration in which the bifacial sheet is in a foldable state and a second configuration in which the bifacial sheet is in a folded state, wherein the bifacial sheet has an upper surface and an inner surface, and is substantially equally divided into 4 foldable quadrants by 3 parallel crease lines in the bifacial sheet, the foldable quadrants including: a first central quadrant and a second central quadrant mutually adjacent to a central crease line in the bifacial sheet, wherein the second central quadrant contains an aperture in the bifacial sheet to allow for deposition of a blood specimen therethrough,

a first outer quadrant adjacent to the first central quadrant, and a second outer quadrant adjacent to the second central quadrant;

(b) the analytic membrane assembly to being disposed on the bifacial sheet, such that the separator membrane is adhered to the upper surface of the second central quadrant and covers the aperture therein and the capture membrane is adhered to the upper surface of the first central quadrant; and

[0090] In certain embodiments, the analytical membrane array may be incorporated in a vertical flow separator device that comprises an elongated base that forms a continuous protective surface around the device housing the analytical membrane array.

[0091] The separator device according to embodiments of the invention may therefore be a vertical flow plasma separation device and may contain therein a removable desiccant paper near the separator membrane and/or the support membrane. In additional embodiments, the vertical flow plasma separation device may have a bifacial sheet having a folded state according to the second configuration of the support assembly that includes the first outer quadrant folded onto the first central quadrant so that the upper surface of the first outer quadrant is placed over the upper surface of the first central quadrant and sealed in place by the adhesive strip affixed to the upper surface of the bifacial sheet along the outer perimeter of the first central quadrant; and the second outer quadrant folded onto the second central quadrant so that the upper surface of the second outer quadrant is placed over the upper surface of the second central quadrant and sealed in place by the adhesive strip affixed to the upper surface of the bifacial sheet along the outer perimeter of the second central quadrant.

[0092] In certain embodiments, the vertical flow plasma separation device may contain the bifacial sheet having the folded state according to the second configuration of the support assembly folded onto the second central quadrant along the central crease line so as to provide the blood separation device in a closed state, the lower surface of the first outer quadrant that is already folded over the first central quadrant being placed over the lower surface of the second outer quadrant that is already folded over the second central quadrant, whereby the lower surface of each of the first and second central quadrants constitutes an outer surface of the vertical flow plasma separation device in a closed state.

[0093] Referring to the drawings, and initially to FIGS. 2-12, a first embodiment of a blood/plasma separation device is generally indicated by (10). The blood/plasma separation device is an elongated bifacial sheet (10) that is preferably self-contained and constructed so as to process a whole blood sample that may be obtained from a finger prick. The blood/plasma separation device according to embodiments may be in a foldable state (i.e., unfolded) as shown in FIGS. 2 and 12, or in a completely folded state as shown in FIG. 11 , including various partially folded states as shown in FIGS. 4-10 and 15-20 and nearly folded states as shown in FIGS. 27 and 28.

[0094] In general, the blood/plasma separation device contains at least the following features: a plurality of foldable quadrants (22, 24, 26, 28) divided by parallel crease lines (41, 42, 43) in the bifacial sheet (10); an aperture (30) in the bifacial sheet used to collect a biological sample (e.g., a blood specimen); and the analytical membrane array disclosed herein, which includes at least a separation membrane (50) and a capture membrane (70).

[0095] As shown in the drawings, the bifacial sheet (10) is preferably elongated and has a substantially uniform width that is substantially less than its longitudinal length. The bifacial sheet (10) can be (but is not limited to) conventional card stock that is preferably water repellent. FIG. 2, shows an embodiment wherein the bifacial sheet (10) is in a flat, but foldable, configuration, and is substantially equally divided into 4 foldable quadrants (22, 24, 26, 28) by three parallel crease lines (41, 42, 43) in the bifacial sheet (10), the foldable quadrants including a first central quadrant (24) and a second central quadrant (26) mutually adjacent to a central crease line (42) in the bifacial sheet, wherein the second central quadrant (26) contains an aperture (30) in the bifacial sheet to allow for deposition of a blood specimen therethrough. The bifacial sheet (10) also includes a first quadrant (22) adjacent to the first central quadrant (24), and a second outer quadrant (28) adjacent to the second central quadrant (26). The analytic membrane assembly is disposed on the bifacial sheet (10) such that the separation membrane (50) is adhered to an upper surface of the second central quadrant (26) and covers the aperture (30) therein, and the capture membrane (70) is adhered to the upper surface of the first central quadrant (24). An adhesive strip (35) may be affixed to the upper surface of the bifacial sheet (10) along an outer perimeter of each of the first central quadrant (24) and the second central quadrant (26) so as to place and optionally seal the upper surface of the first outer quadrant (22) directly over the upper surface of the first central quadrant (24) to enclose and protect the capture membrane (70) located on the first central quadrant (24). The second outer quadrant (28) is also foldable onto the second central quadrant (26) so as to place and optionally seal the upper surface of the second outer quadrant (28) directly over the upper surface of the second central quadrant (26) to enclose and protect the separation membrane (50) comprised on the second central quadrant (26).

[0096] As shown in FIG. 12, the vertical flow blood/plasma separation device may also optionally contain a removable desiccant paper (100) near the separator membrane (50) and/or the collection membrane (70). As shown in FIG. 12, the adhesive strip(s) (35) may be provided on opposing rails (90).

[0097] In certain embodiments, the separator device is a lateral flow blood separation device. In such embodiments, the separator device comprises:

(a) a support assembly of two sheets separably coupled together in a foldable configuration and in a folded configuration, the sheets including an elongated bifacial base sheet having an upper surface and an inner surface, and substantially equally divided into a first and second foldable panel by a central folding line, the first foldable panel being further divided into an outer quadrant and an inner quadrant that is between the outer quadrant of the first panel and the second panel, and an elongated bifacial cover sheet having an upper surface and a lower surface, a size corresponding to that of the first panel of the base sheet, and an aperture to allow for deposition of a blood specimen therethrough;

[0098] The lateral flow plasma separation device is furthermore separable from the capture membrane by adhesion to the lower surface of the cover sheet, whereby the separator membrane becomes adhered to the lower surface of the cover sheet placed thereon and separated from the capture membrane as a result of the cover sheet folding onto the second panel of the base sheet; and the outer quadrant is foldable onto the central quadrant so as to place and optionally seal the upper surface of the outer quadrant directly over the upper surface of the central quadrant to enclose and protect the capture membrane comprised on the central quadrant.

[0099] FIGS. 13-23 relate to a lateral flow blood/plasma separator device according to alternative embodiments of the invention. Specifically, the analytic membrane array (FIG. 14) used in such embodiments is configured for longitudinal flow. In general, the lateral flow blood/plasma separator device contains at least the following features: a plurality of foldable quadrants (144, 146, 148) divided by parallel crease lines (41, 43) in the bifacial sheet (10); and aperture (30) in an upper bifacial sheet (110) used to collect a biological sample (e.g., a blood specimen); and the analytical membrane array disclosed herein, which includes at least a separation membrane (50), a capture membrane (70), and a separator backing (80). [00100] As shown in FIG. 23, the lateral flow blood/plasma separator device may also optionally include a removable desiccant paper (100) near the separator membrane (50) and/or the collection membrane (70).

[00101] FIG. 24 shows a multi-spot blood/plasma separator device (150) according to certain embodiments, wherein whole blood specimens may be stored in a left quadrant (152) and plasma specimens may be stored in a right quadrant (154), wherein the quadrants separated and foldable by a central crease line (155). The multi-spot blood/plasma separator device (150) may additionally include adhesive strip(s) (35) on opposing rails (90) and tabs (95, 97) extending from each quadrant at opposite sides of the respective quadrants so as to enable ease of handling (closing and opening) of the device. As shown by the configuration exemplified in FIG. 25, the multi-spot blood/plasma separator device according to may comprise a 2 x 2 plasma card (shown by right quadrant (154)), and optionally also a 2 x 2 whole blood card (shown by left quadrant (152)), wherein the cards each provide for two rows of two spots for deposition and storage of the plasma and whole blood specimens, respectively. The whole blood spots for deposition and storage of whole blood may furthermore include a slightly raised circular perimeter (160) to facilitate the determination that an appropriate blood volume has been applied to the device. Another exemplified configuration of the blood/plasma separator device of the invention is shown in FIG. 25, wherein each quadrant (152, 154) contains one row of four spots (1 x 4) for deposition and storage of the plasma and/or whole blood specimens.

[00102] FIG. 26 shows yet another exemplified configuration of the blood/plasma separator device, which includes a 2 x 1 plasma card (two rows of one spot each) for deposition and storage of plasma, as well as a separate (independent) quadrant (170) for deposition and storage of additional whole blood specimens for further analysis. In FIG. 27, a blood/plasma separator device according to a similar embodiment of the invention is shown in a partially folded configuration, wherein tabs (95, 97) extend from each quadrant at opposite sides of the respective quadrants so as to enable ease of handling (closing and opening) of the device, and a separate (independent) quadrant (180) is provided for deposition and storage of plasma specimens for further analysis.

[00103] According to embodiments of the invention, the blood/plasma separator device containing the analytical membrane assembly and/or plasma card as described herein may have, in an unfolded configuration, a length of from about 50 mm to 250 mm (as measured from edge to edge and not including any of the optional tabs (95, 97) that are further described herein for use in various embodiments) and a width of from about 20 mm to about 60 mm. In specific embodiments, the blood/plasma separator device containing the analytical membrane assembly and/or plasma card as described herein may have an unfolded length of from about 100 mm to about 200 mm, and a width of from about 40 mm to about 50 mm. For example, a blood/plasma separator device having a 1 x 4 configuration (each foldable quadrant having one row of four spots) as shown in FIG. 25 may have: a length of about 200 mm, or about 100 mm if in a folded (i.e., closed) configuration; and a width of about 40 mm. Alternatively, a blood/plasma separator device having a 2 x 2 configuration (each foldable quadrant having two rows of two spots) as shown in FIG. 24 may have a length of about 100 mm, or about 50 mm if a folded configuration; and a width of about 50 mm.

[00104] In embodiments, the analytical membrane array and/or the separator device that may contain the analytical membrane array, are compatible for use with conventional handheld scanners and mobile/cellular smart phones. In certain embodiments, the separator device may be a hand held point-of-care plasma separator.

[00105] According to embodiments of the invention, personal identifying information is furthermore obtained from each patient providing a biological specimen. Such personal identifying information may include, e.g., name, sex, date of birth, ethnic background, location of testing, etc. (if relevant to the tests performed). If certain types of genetic markers are to be tested for a patient, that information may also be obtained and provided. Thus, according to the described methods, personal information that identifies the patient is additionally associated with each corresponding specimen.

[00106] As shown in FIG. 28, a blood/plasma separation device according to a preferred embodiment of the invention additionally includes identification information or technology (120) on an outer surface thereof. For example, the identifying information or technology may be integrated into an outer surface of the bifacial sheet (10). Any suitable RFID chip or similar identification technology may be integrated onto the cardstock of the blood/plasma separator device according to embodiments of the invention, including, e.g., single-dimensional (ID) barcodes, two-dimensional (2D) barcodes, QR code, etc. FIGS. 29A-29C depict various identifying information or technology formats suitable for use in exemplified embodiments of the invention. [00107] The identifying technology (e.g., barcode) provided on an outer surface of a device according to embodiments of the invention may contain information specific to the patient associated with the biological specimen stored therein, including variable personal information (such as address, signature, birthdate, etc.), and/or biometric information (e.g., a fingerprint, a facial image or template. To protect the information that is provided on a label or by the technology on an outer surface of the device, a layer of overlaminate may additionally be provided over the label or technology. Material suitable for forming such protective layers are known to those skilled in the art of making identification documents and any of the conventional materials may be used provided they have sufficient transparency. Furthermore, the label or identifying technology may be provided on the outer surface of the device of the invention in any desired size and conventional thicknesses. .

[00108] According to embodiments of the invention, the personal identifying information is presented and stored in a way that respects the privacy of the patient in compliance with the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"). Various methods and systems for protecting personal identifying information of patients are known in the art, and may be employed in practicing embodiments of the invention. For example, an image relating to protected health information may be electronically modified so as to delete patient identifying information, or privacy criteria may be provided that restrict and control access to the personal identifying information. See, e.g., U.S. Patent No. 7,936,913 (describing a hematology imaging system capable of selectively removing sensitive patient identifying information from an image or including the same in the image); U.S. Patent No. 7,992,002 (relating to an EMR data depository with an access controller for approving or denying a request for access to data); U.S. Patent No. 8,019,620 (disclosing a system including request manager, release manager, and privacy status manager modules for validating information requests according to privacy criteria associated with a corresponding patient). The full disclosures of the above- referenced patents are incorporated by reference herein.

[00109] In embodiments, the identification technology may be in a format to allow for cloud-based transmission and storage of the personal identifying information of the associated patient by methods and systems known in the art. Identification technology suitable for use in embodiments of the invention may be accessed via any type and/or form of cloud services or systems to provide a cloud-based information exchange and, optionally, authentication over one or more networks. The network and/or network topology may be selected from any network or network topology known to those skilled in the art. Identification technology for cloud-based information transmission may comprise any combination of hardware and software and may be deployed as an application with software installed on one or more devices. Suitable applications for use with embodiments of the invention may process and store various types and forms of patient records.

[00110] An optional authentication feature of the identification technology may be particularly useful in resource-limited settings. The authentication feature is preferably designed to allow for authentication of stored samples in the associated device by a smart phone application. For example, according to the exemplified embodiment shown in FIG. 30, results of authentication of the biological specimen(s) of the analytic membrane are shown inside the circled portion of FIG. 30A (unchanged background being shown in red greyscales and unchanged foreground shown in green greyscales). Any suspected alterations are localized as "hot spots," as shown inside the circled portion of FIG. 30B. According to the exemplified authentication system, a fake or fraudulent barcode or identification technology will not result in transmission of information, as shown in the empty circled area of FIG. 30C. The application interface results depicted in FIG. 30 were generated using TRAXSECUR; however, other security and authentication smart phone applications may be used in practicing the invention.

[00111] Biological specimens suitable for use in the analytical membrane array or a device containing the same includes, but are not limited to, whole blood, plasma, urine, saliva, sputum, semen, vaginal lavage, bone marrow, cerebrospinal fluid, other physiological or pathological body liquids, or any of the combinations thereof. In certain embodiments, the biological specimen is human body fluid, such as whole blood containing the analytes of interest, such as proviral DNAs and/or other nucleic acids (including either or both DNA and RNA molecules), and/or plasma proteins, such as, e.g., Troponin, monoclonal kappa and lambda free light chains, Cystatin C and Carbohydrate- Deficient Transferrin (CDT). In certain embodiments, the analytes of interest are nucleic acids and the biological specimens comprise at least 5 ng to 1 μg of either or both DNA or RNA molecules. In yet other embodiments, the biological specimen is contained in liquid suspension that may include (but is not limited to), e.g., cell suspension, liquid extracts, tissue homogenates, media from DNA or RNA synthesis, saline, or any combinations thereof.

[00112] In certain embodiments, the biological specimen administered to the analytical membrane array is whole blood. In such embodiments, a liquid suspension of the whole blood sample is absorbed and dried on the separation membrane, where solid components of whole blood are captured, and the fluid component of whole blood is drawn through gravity and capillary action onto the capture membrane, where the cell- free plasma is captured. In these embodiments, the first portion of the analytical membrane array, the separation membrane, filters blood components and allows for flow of the fluid component through release of serum/plasma. Embodiments of the invention provide that when whole blood is loaded onto the separation membrane, the separation membrane separates the solid components from the fluid component of whole blood by capturing the solid components (e.g., WBCs, RBCs, platelets, and/or other cellular components), whereby the fluid component of the whole blood can be drawn onto the capture membrane through gravity and capillary action so as to separate serum and/or plasma from the whole blood. The separation membrane and the capture membrane can be physically separated and removed for independent assays and analysis of their respective filtered components.

[00113] In certain embodiments, the cell-free plasma captured on the capture membrane of the analytical membrane array may be further reconstituted in a reconstitution media and then removed and recovered. The recovered cell-free plasma contains an analyte of interest, for instance, nucleic acids of interest including but not limited to DNA and RNA, which can be used for viral load quantitation, genotyping, drug resistance testing, or other analysis of a viral nucleic acid of interest.

[00114] Embodiments of the invention further provide for subsequent analysis of the analytes of interest contained in the biological specimen that are recovered from each portion (or membrane) of the device into the reconstitution medium, such as molecular- grade water. As used herein, the term "subsequent analysis" includes any analysis which may be performed on recovered biological specimens stored in reconstitution medium. Alternatively, the analytes of interest contained in the biological specimen may be isolated, purified or extracted prior to analysis using methods known in the art. The analytes of interest may be subjected to chemical, biochemical or biological analysis. In one of the preferred embodiments, the analytes of interest are nucleic acids including either or both proviral and/or viral DNA or RNA molecules that can be detected or analyzed with or without prior extraction, purification or isolation. DNA or RNA extraction, purification or isolation, if necessary, is performed based on methods known in the art. Examples of subsequent analysis include MS/MS, "Tandem Mass Spectrometry," LC/MS, LC/MS/MS, MALDI-TOF, TOF, GC-MS, ESI ElectroSpray Ionization, FAB Fast Atom Bombardment (occasionally referred to as SIMS or Secondary Ion Mass Spectrometry), MS Fourier Transform Mass Spectrometry, IRMPD InfraRed Multi- Photon Dissociation, LSIMS Liquid Secondary Ion Mass Spectrometry (synonymous with FAB), MALDI Matrix Assisted Laser Desorption Ionization, NRMS Neutralization- Reionization Mass Spectrometry, REMPI Resonance Enhanced Multi- Photon Ionization, SORI-CAD Sustained Off-Resonance Irradiation- Collisionally Activated, wGC-MS Gas Chromatography-Mass Spectrometry, HR-MS High Resolution Mass Spectrometry, IRMPD INfraRed Multi-Photon Dissociation, LSIMS Liuie dSecnary Ion Masspolymerase chain reaction (PCR), ligase chain reaction (LCR), reverse transcriptase initiated PCR, DNA or RNA hybridization techniques including restriction fragment length polymorphism (RFLP), viral DNA or RNA detection and quantification, viral load tests, DNA or RNA genotyping, etc. "Subsequent analysis" also includes other techniques using genetic probes, genomic sequencing, enzymatic assays, affinity labeling, methods of detection using labels or antibodies and other similar methods. In certain embodiments, the subsequent results of the analyses of reconstituted and recovered analytes from each portion of the array (i.e., from each membrane) can be evaluated together for more precise detection and diagnosis. For instance, inventive methods of using the analytic array include for HIV detection by detecting both the proviral DNAs isolated from the white cells of the whole blood absorbed on the separator membrane, as well as the viral DNAs or RNAs in plasma reconstituted and recovered from the capture membrane from the same whole blood specimen.

[00115] In embodiments, analytes of interest include, but are not limited to, nucleic acids, proteins, carbohydrates, lipids, whole cells, cellular fragments, a whole virus or viral fragments. In certain embodiments, the analytes of interest are nucleic acids including either or both DNA and RNA molecules. Certain embodiments provide for improved systems and methods for the detection and quantification of RNA, e.g., whole virus for determining viral load and genotyping in a biological specimen or subject. [00116] In certain embodiments, a nucleic acid of interest may be HCV or other single stranded RNA viruses. In certain embodiments, the nucleic acid of interest is HIV or other retroviruses. In certain embodiments, the nucleic acid of interest is HBV or other double stranded DNA viruses. In certain embodiments, the nucleic acid of interest is Influenza or other double stranded RNA viruses. In certain embodiments, the nucleic acid of interest is Parvovirus B19 or other single stranded DNA viruses. In certain embodiments, the nucleic acid of interest is contained within the HCV genome or the genome of other single stranded RNA viruses. In certain embodiments, the nucleic acid of interest is contained within the HIV genome or the genome of other retrovirus. In certain embodiments, the nucleic acid of interest is HBV genome or the genome of other double stranded DNA viruses. In certain embodiments, the nucleic acid of interest is Influenza genome or the genome of other double stranded RNA viruses. In certain embodiments, the nucleic acid of interest is Parvovirus B19 or the genome of other single stranded DNA virus.

[00117] In further embodiments, the invention relates to subsequent analysis using a recovered biological specimen that contains analytes of interest. The analytes of interest may be proviral DNA and/or RNA molecules that are detected or analyzed using analytical and diagnostic methods known in the art. In some embodiments, the analytes of interest may be white blood cells containing proviral DNAs and/or intact virus, such as HCV or HIV, and the biological specimen recovered from the device is used for evaluation and analytical measurements with reproducibility, accuracy, and precision.

[00118] The dual stage analytical membrane array, a device containing the analytical membrane array, and methods of using the same allow for biological testing of air-dried bodily fluid samples without the need for refrigerated or frozen shipping and storage. The inventive analytical membrane array, device, and methods provide the capability to significantly reduce the costs of shipping infectious materials worldwide, especially those associated with large clinical trials. Moreover, the inventive analytical assembly array, device and methods for preserving biological specimens are applicable to and include a wide range of esoteric and standard clinical testing, including qualitative and quantitative nucleic acid analysis.

[00119] The analytical membrane array has an ability to absorb a liquid suspension readily and quickly, as well as to release the biological specimen containing analytes of interest consistently, efficiently, and precisely. In these embodiments, the separator membrane provides for filtering of blood components and serum/plasma separation. According to embodiments, when a biological specimen, such as the whole blood, is loaded onto the first portion of the asymmetric absorbent matrix of the invention, the primary stage with the asymmetric matrix can capture the solid components (e.g., WBCs, RBCs, and/or other cellular components of whole blood), and the fluid component of the biological specimen can be drawn onto the second stage with the polyolefin matrix through gravity and capillary action so as to separate the components in the specimens (e.g., serum and/or plasma from the whole blood). Prior to reconstitution and recovery of the components captured in either matrix, respectively, the two matrices can be is physically removed and/or broken off.

[00120] Methods according to embodiments further also may include an intermediate step of applying a stabilizing composition to the analytical membrane array to protect the analytes of interest against degradation. Depending upon the analytes of interest, the stabilizing composition, as discussed above, may include but is not limit to one or more of a weak base, a chelating agent, a protein denaturing agent such as a detergent or surfactant, a nuclease inhibitor, and a free radical trap. Particularly for protection of unstable RNA, the stabilizing composition may include RNase inhibitors and inactivators, genetic probes, complementary DNA or RNA (or functionally equivalent compounds), proteins and organic moieties that stabilize RNA or prevent its degradation.

[00121] According to certain embodiments, also provided is a method for recovering from each membrane of the analytical membrane array in the compression device the captured components from the biological specimen containing analytes of interest. In certain embodiments, the method includes the following steps: a) applying reconstitution medium to each portion of the analytical membrane array to rehydrate the bound biological specimen containing analytes of interest, and b) compressing the membrane to release a portion of the biological specimen. According to the present invention, the reconstitution medium is molecular-grade water. In other embodiments, the reconstitution medium includes the components of IX phosphate buffered saline (PBS) or nuclease-free water optionally with the addition of sodium azide or other antimicrobial agent. The reconstitution medium may also include any number or combinations of available biological preservatives or blood anticoagulants including but not limited to ethylenediaminetetraacetic acid (EDTA), sodium citrate, and heparin. PBS or nuclease- free water serves as the sterile and neutral medium for the rehydration, re-suspension, and recovery of the analyte(s) of interest from the membrane array. When included, antimicrobial agents such as sodium azide prevent microbial growth and subsequent contamination with RNases. When included, biological preservatives such as EDTA, sodium citrate, and heparin serve as anticoagulants and or chelating agents.

[00122] In certain embodiments, the analytical membrane assembly according to the test data shows a favorable rating, based on an average 0.05 ml, 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml, 0.6 ml, 0.7 ml, 0.8 ml, or 0.9 ml, 1.0 ml, 1.5 ml, 2.0 ml, 2.5 ml, 3.0 ml, or greater, sample of a liquid suspension of a biological specimen containing an analyte of interest.

[00123] The volume of each membrane of the analytical membrane assembly may or may not expand upon absorption of the liquid suspension, and may or may not contract upon drying. However, a liquid saturated membrane can be compressed to release entrained fluid containing analyte, due to its porosity, by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, or more of its saturated volume. Volumetric compression is one convenient technique for release of the reconstituted biological specimen, however, any other means, such as centrifugation or vacuum pressure, can alternatively be employed to release the biological specimen from the array.

[00124] A plasma/blood separation device according to embodiments described herein may optionally include a desiccant, either a natural or synthetic desiccant, inside the container to maintain the dried state of the membrane and integrity of the analytes of interest on the membrane within the enclosed container. In certain embodiments, the desiccant is in vaporous communication with the analytical membrane array in the compression device having a dye indicator reactive with moisture whereby the desiccant changes to a bright color when exposed to humidity or moisture. In certain embodiments, the desiccant is in vaporous communication with the analytic membrane assembly that an air permeable barrier is formed in-between the desiccant and the assembly inside the device. The desiccant used in the device is commonly known in the art, including but not limited to montmorillonite clay, lithium chloride, activated alumina, alkali alumino- silicate, DQ11 Briquettes, silica gel, molecular sieve, calcium sulfate, and calcium oxide. The desiccant may also have been provided with a colorimetric indicator of water content. In other embodiments, a desiccant may not be needed inside the device containing the analytic membrane assembly.

[00125] The analytical membrane array may optionally include a composition absorbed to a surface thereof, wherein the composition protects against degradation of the analytes of interest contained in the biological specimens. As used herein, the term "protects against degradation of the analytes of interest" means that a membrane assembly in the device of the invention maintains the stored analytes of interest contained in the biological specimens in a substantially nondegraded form, providing that the analytes of interest are suitable for many different types of subsequent analytical procedures. Protection against degradation may include protection against substantial damaging of analytes of interest caused by chemical or biological agents including action of bacteria, free radicals, nucleases, ultraviolet radiation, oxidizing agent, alkylating agents, or acidic agents (e.g., pollutants in the atmosphere). In certain embodiments, the composition absorbed on the analytical assembly array (i.e., on a surface of a membrane thereof) may include one or more of a weak base, a chelating agent, a protein denaturing agent such as a detergent or surfactant, a nuclease inhibitor, and a free radical trap. In a case where the stored analyte of interest is RNA, particularly unstable RNA, the composition may include RNase inhibitors and inactivators, genetic probes, complementary DNA or RNA (or functionally equivalent compounds), proteins and organic moieties that stabilize RNA or prevent its degradation.

[00126] Another composition which protects against degradation which may be optionally used is an oxygen scavenger element. Suitable oxygen scavenging elements are well-known to those skilled in the art. Non-limiting examples of oxygen scavenging elements include but are not limited to compositions comprising metal particulates reactive with oxygen such as transition metals selected from the first, second or third transition series of the periodic table of the elements, and include manganese II or III, iron II or III, cobalt II or III, nickel II or III, copper I or II, rhodium II, III or IV, and ruthenium. The transition metal is preferably iron, nickel or copper. An example of an iron oxygen scavenging element is D500 from Multisorb. Other commercially available oxygen scavengers may also be purchased from companies such as Mitsubishi, Dow, or the like. Other examples of oxygen scavenging element may be enzymes which consumes, depletes or reduces the amount of oxygen from the given environment without negatively affecting the samples of interests.

[00127] In other embodiments, the compression device may optionally comprise a modified atmosphere such as nitrogen or argon through a well-known gas purging process prior to sealing, shipping, or storing. The term "modified atmosphere" refers to any replacing or altering normal atmospheric gas compositions with at least one inert gas or gas which does not degrade the sample of interests.

[00128] As used herein, a "weak base" suitable for the composition of the invention may be a Lewis base which has a pH of about 6 to 10, preferably about pH 8 to 9.5. The weak base suitable for the composition of the invention may, in conjunction with other components of the composition, provide a composition pH of 6 to 10, preferably, about pH 8.0 to 9.5. Suitable weak bases according to the invention include organic and inorganic bases. Suitable inorganic weak bases include, for example, an alkali metal carbonate, bicarbonate, phosphate or borate (e.g., sodium, lithium, or potassium carbonate). Suitable organic weak bases include, for example, tris-hydroxymethyl amino methane (Tris), ethanolamine, triethanolamine and glycine and alkaline salts of organic acids (e.g., trisodium citrate). A preferred organic weak base is a weak monovalent organic base, for example, Tris. The weak base may be either a free base or a salt, for example, a carbonate salt. It is believed that the weak base may provide a variety of functions, such as protecting the analytes of interest from degradation, providing a buffer system, ensuring proper action of the chelating agent in binding metal ions, and preventing the action of acid nucleases which may not be completely dependent on divalent metal ions for functioning.

[00129] As used herein, a "chelating agent" is any compound capable of complexing multivalent ions including Group II and Group III multivalent metal ions and transition metal ions (e.g., Cu, Fe, Zn, Mn, etc.). In certain embodiments, the chelating agent is ethylene diamine tetraacetic acid (EDTA), citrate or oxalate. It is believed that one function of the chelating agent is to bind multivalent ions which if present with the stored biological specimen may cause damage to the analytes of interest, especially to nucleic acids. Ions which may be chelated by the chelating agent include multivalent active metal ions, for example, magnesium and calcium, and transition metal ions, for example, iron. Both calcium and magnesium are known to promote nucleic acid degradation by acting as co-factors for enzymes which may destroy nucleic acids (e.g., most known nucleases). In addition, transition metal ions, such as iron, may readily undergo oxidation and reduction and damage nucleic acids by the production of free radicals or by direct oxidation.

[00130] The composition can further include a protein denaturing agent in the second stage where the analytes of interest are nucleic acids. As used herein, a "protein denaturing agent" functions to denature non-nucleic acids compounds, for example, nucleases. If the protein denaturing agent is a detergent or a surfactant, the surfactant may also act as a wetting agent to facilitate the uptake of a sample by the dry solid capture membrane. The terms "surfactant" and "detergent" are synonymous and may be used interchangeably throughout the specification. Any agent that denatures proteins without substantially affecting the nucleic acids of interest may be suitable for the invention. In certain embodiments, protein denaturing agents include detergents. As used herein "detergents" include ionic detergents, preferably anionic detergents. An anionic detergent suitable for the invention may have a hydrocarbon moiety, such as an aliphatic or aromatic moiety, and one or more anionic groups. Particularly, suitable anionic detergents include sodium dodecyl sulphate (SDS) and sodium lauryl sarcosinate (SLS). The ionic detergent causes inactivation of a microorganism which has protein or lipid in its outer membranes or capsids, for example, fungi, bacteria or viruses. This includes microorganisms which may be pathogenic to humans or which may cause degradation of nucleic acids. It is believed that inactivation of a microorganism by a detergent is a result of destruction of the secondary structure of the organisms external proteins, internal proteins, protein containing membranes, or any other protein necessary for viability. However, the detergent may not inactivate some forms of organisms, for example, highly resistant bacterial spores and extremely stable enteric virions.

[00131] The composition may optionally include a free radical trap. As used herein, a "free radical trap" is a compound which is sufficiently reactive to be preferred, over a DNA molecule or a component thereof, as a reactant with a free radical, and which is sufficiently stable not to generate damaging free radicals itself. Examples of a suitable free radical trap include: uric acid or a urate salt, mannitol, benzoate (Na, K, Li or tris salt), 1 -3 dimethyl uric acid, guanidine, guanine, thymine, adenine, cytosine, in N-acetyl-histidine, histidine, deferoxamine, dimethyl sulfoxide, 5 '5' dimethyl pyrroline-N-oxide, thiocyanate salt and thiourea. Suitable free radical traps include mannitol, thiocyanate salts, uric acid or a urate salt. It is believed that the longer the period of time for which the nucleic acid is to be stored the more likely that a free radical trap may be advantageously included in the composition absorbed to the solid analytical membrane array. Even if the nucleic acid is only to be stored for a matter of minutes, a free radical trap may still be incorporated into the composition. It is believed that one function of the free radical trap may be to trap nucleic acid damaging free radicals. For example, when the free radical trap used is uric acid or urate salt it may be converted to allantoin which may also act as a free radical trap that accepts free radicals that would otherwise damage nucleotide bases, for example, guanine. In certain embodiments, the free radical trap reacts with free radicals regardless of source (including free radicals present in the air). Free radicals may be generated through oxidation or reduction of iron in biological specimen, such as blood. Typically, free radicals are believed to be generated by spontaneous oxidation of the groups which are present, for example, in denatured serum protein of blood. Free radicals may also be generated by radiation such as UV light, x-rays and high-energy particles. In addition, free radical traps which are also a weak acid, e.g. uric acid, can also function as a component of the buffering system provided by the weak base discussed above. Also, the free radical trap may enhance removal of a stored sample of nucleic acids if in situ processing is not desired.

[00132] In embodiments, time periods for which biological specimen may be preserved may be as short as the time necessary to transfer a sample of biological specimen from a collection source to the place where subsequent analysis is to be performed. Therefore, such preservation may be for a period of several minutes, hours, days, months, or even greater.

[00133] Temperature conditions under which a biological specimen may be stored in a plasma separation device according to various embodiments described herein is not limited. Typically, samples of biological material are shipped and/or stored at ambient or room temperature, for example, from about 15°C to about 40°C, preferably from about 15°C to about 25 °C. In another embodiment the samples may be stored in a cool environment. For example, in short- term storage, the samples can be refrigerated at about 2°C to about 10°C. In yet another example, the samples may be refrigerated at about 4°C to about 8°C. In another example, in long-term storage, the samples can be frozen at about -80°C to about -10°C. In yet another example, the samples can be frozen from about -60°C to about -20°C. In addition, the device may preferably but not necessarily be stored in dry or desiccated conditions or under an inert atmosphere.

[00134] It should be understood that the foregoing disclosure relates to certain embodiments of the present invention and that various changes or modifications may be made therein without departing from the scope of the invention. The invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope thereof. On the contrary, it is to be clearly understood that resort may be had to various other embodiments, modifications, and equivalents thereof, which, after reading the description herein, may suggest themselves to those skilled in the art without departing from the spirit of the present invention and/or the scope of the appended claims. It is, therefore, intended that the appended claims cover all such equivalent variations as fall within the true spirit and scope of the invention.

[00135] The detailed description set forth above is provided to aid those skilled in the art in practicing the invention. However, the invention described and claimed herein is to be limited in scope by the specific embodiments described above, as these embodiments are presented as mere illustrations of several aspects of the invention. Any combinations and modifications of the described methods and components, and compositions used in the practice of the methods, in addition to those not specifically described, will become apparent to those skilled in the art based on the present disclosure and do not depart from the spirit or scope of the present invention. Such variations, modifications, and combinations are also encompassed by the present disclosure and fall within the scope of the appended claims.

[00136] Various embodiments discussed herein will be further understood by way of the experimental results of comparative studies of various separation materials that are provided in Table 1 below.

Claims

WHAT IS CLAIMED IS:

1. An analytic membrane array comprising:

a planar asymmetric separator membrane of a polysulfone polymer material having a porosity that gradually decreases from an upstream side to a downstream side of the membrane so as to filter and trap solid components of a biological specimen deposited on the separator membrane; and

a planar capture membrane made of a substantially hydrophobic polyolefin material or cotton or cellulose that allows for flow of a liquid component of the biological specimen therethrough, wherein the capture membrane dissociably overlaps with at least a portion of the separator membrane at a downstream side of the separator membrane so as to provide for vertical or lateral downstream flow of the liquid component of the biological specimen from the separator membrane to the capture membrane for further filtering.

2. The analytic membrane array of claim 1 , wherein the separator membrane is configured to filter and trap solid components of a biological specimen, the biological specimen being selected from the group consisting of whole blood, plasma, urine, saliva, sputum, semen, vaginal lavages, bone marrow and cerebrospinal fluid.

3. The analytic membrane array of claim 1 , wherein the separator membrane is configured to filter and trap solid components of a whole blood specimen and the capture membrane is configured to separately filter and trap a plasma fraction or filtrate of the whole blood specimen.

4. The analytic membrane array of claim 1 , wherein a ratio of a surface area of the separator membrane to a surface area of the capture membrane is at least 2:1.

5. The analytic membrane array of claim 1 , wherein a size and/or shape of the separator membrane is the same as a size and/or shape of the capture membrane.

6. The analytic membrane array of claim 1 , wherein the separator membrane has a pore size ranging from 0.1 -20 μ m.

7. The analytic membrane array of claim 1 , wherein the separator membrane and/or the capture membrane has a substantially circular shape.

8. The analytic membrane array of claim 1 , wherein the separator membrane has a teardrop shape with an elongated portion near the downstream side thereof, and the elongated portion of the separator membrane separably overlaps with at least a portion of the capture membrane.

9. The analytic membrane array of claim 1 , wherein the polysulfone polymer material of the separator membrane is at least one polysulfone polymer or matrix selected from the group consisting of asymmetric sub-micron polysulfone and asymmetric super micron polysulfone.

10. The analytic membrane array of claim 1 , wherein the substantially hydrophobic polyolefin material of the capture membrane comprises a plurality of fibers of polypropylene coated with hydrophobic polyethylene.

11. The analytic membrane array of claim 1 , wherein the separator membrane and/or the capture membrane comprises microglass fibers.

12. A vertical flow blood separation device, comprising:

(a) a support assembly of an elongated bifacial sheet having a first configuration in which the bifacial sheet is in a foldable state and a second configuration in which the bifacial sheet is in a folded state, wherein the bifacial sheet has an upper surface and an inner surface, and is substantially equally divided into 4 foldable quadrants by 3 parallel crease lines in the bifacial sheet, the foldable quadrants including:

a first central quadrant and a second central quadrant mutually adjacent to a central crease line in the bifacial sheet, wherein the second central quadrant contains an aperture in the bifacial sheet to allow for deposition of a blood specimen therethrough,

(b) the analytic membrane assembly of claim 1 disposed on the bifacial sheet, such that the separator membrane is adhered to the upper surface of the second central quadrant and covers the aperture therein and the capture membrane is adhered to the upper surface of the first central quadrant; and

(c) an adhesive strip affixed to the rails of the upper surface of the bifacial sheet along an outer perimeter of each of the first central quadrant and the second central quadrant, wherein the first outer quadrant is foldable onto the first central quadrant so as to place and optionally seal the upper surface of the first outer quadrant directly over the upper surface of the first central quadrant to enclose and protect the capture membrane located on the first central quadrant, and the second outer quadrant is foldable onto the second central quadrant so as to place and optionally seal the upper surface of the second outer quadrant directly over the upper surface of the second central quadrant to enclose and protect the separator membrane comprised on the second central quadrant.

13. The vertical flow blood separation device of claim 12, further comprising a removable desiccant paper near the separator membrane and/or the support membrane.

14. The vertical flow blood separation device according to claim 12, wherein:

the second central quadrant is foldable along the central crease line onto the first central quadrant so as to separably place the upper surface of the second central quadrant and the second outer quadrant over the upper surface of the first central quadrant and the first outer quadrant, respectively, and to allow for the capture membrane to separably overlap with at least a portion of the separator membrane so that, upon application of a blood specimen through the aperture in the second central quadrant, the downstream flow of the liquid component of the blood specimen from the separator membrane to the capture membrane proceeds in a vertical direction;

the first outer quadrant is foldable onto the first central quadrant so as to separably place the upper surface of the first outer quadrant over the upper surface of the first central quadrant; and

the second outer quadrant is foldable onto the second central quadrant so as to separably place the inner surface of the second outer quadrant over the inner surface of the second central quadrant.

15. The vertical flow blood separation device of claim 12, wherein the bifacial sheet having a folded state according to the second configuration of the support assembly includes:

the first outer quadrant folded onto the first central quadrant so that the upper surface of the first outer quadrant is placed over the upper surface of the first central quadrant and sealed in place by the adhesive strip affixed to the upper surface of the bifacial sheet along the outer perimeter of the first central quadrant; and

the second outer quadrant folded onto the second central quadrant so that the upper surface of the second outer quadrant is placed over the upper surface of the second central quadrant and sealed in place by the adhesive strip affixed to the upper surface of the bifacial sheet along the outer perimeter of the second central quadrant.

16. The vertical flow blood separation device according to claim 15, wherein the bifacial sheet having the folded state according to the second configuration of the support assembly further includes the first central quadrant folded onto the second central quadrant along the central crease line so as to provide the blood separation device in a closed state, the lower surface of the first outer quadrant that is already folded over the first central quadrant being placed over the lower surface of the second outer quadrant that is already folded over the second central quadrant, whereby the lower surface of each of the first and second central quadrants constitutes an outer surface of the vertical flow blood separation device in a closed state.

17. The vertical flow blood separation device according to claim 16, comprising a label adhered to an outside surface of the blood separation device that is in a closed state, the label including identifying information.

18. A lateral flow blood separation device, comprising:

(a) a support assembly of two sheets separably coupled together in a foldable configuration and in a folded configuration, the sheets including

an elongated bifacial base sheet having an upper surface and an inner surface, and substantially equally divided into a first and second foldable panel by a central folding line, the first foldable panel being further divided into an outer quadrant and an inner quadrant that is between the outer quadrant of the first panel and the second panel, and an elongated bifacial cover sheet having an upper surface and a lower surface, a size corresponding to that of the first panel of the base sheet, and an aperture to allow for deposition of a blood specimen therethrough;

(b) the analytic membrane assembly of claim 1 disposed between the first panel of the base sheet and the cover sheet such that the separator membrane is located atop the outer quadrant of the first panel,

wherein the separator membrane has a teardrop shape with an elongated portion near the downstream side thereof, the elongated portion of the separator membrane separably overlapping with at least a portion of the capture membrane;

19. The lateral flow blood separation device according to claim 18, wherein:

the separator membrane is separable from the capture membrane by adhesion to the lower surface of the cover sheet, whereby the separator membrane is adhered to the lower surface of the cover sheet placed thereon and separated from the capture membrane as a result of the cover sheet folding onto the second panel of the base sheet; and

20. The lateral flow blood separation device according to claim 18, further comprising a removable desiccant paper near the separator membrane and/or the support membrane.

21. A blood/plasma separation device, comprising:

a separator membrane having a porosity that gradually decreases from an upstream side to a downstream side so as to filter and trap solid components of a blood sample deposited on the separator membrane; and

a plasma collection chamber separably adhered to a lower surface of the downstream side of the separation membrane, wherein the plasma collection chamber is configured to collect and store plasma that has been separated from a blood sample by flow from an upstream side to a downstream side of the separator membrane.

22. The blood/plasma separation device according to claim 21 , wherein the plasma collection chamber overlaps with at least a portion of the separator membrane at a downstream side of the separator membrane so as to provide for vertical downstream flow of the liquid plasma component of the whole blood sample from the separator membrane to the plasma collection chamber for storage or further filtering.

23. A blood/plasma separation device, comprising:

(c) a plurality of spots provided on the second quadrant for deposition and storage of plasma specimens of the patient; and (d) an adhesive strip affixed to the upper surface of the bifacial sheet along an outer perimeter of the second quadrant, wherein the first outer quadrant is foldable onto the first central quadrant so as to place and optionally seal the upper surface of the first quadrant directly over the upper surface of the second quadrant to enclose and protect the specimens deposited therein.

24. The blood/plasma separator device according to claim 23, wherein each quadrant comprises a tab extending from opposite end sides thereof so as to enable ease of folding together and subsequently separating the first and second quadrants from each other.

25. The blood/plasma separator device according to claim 23, wherein the plurality of spots provided on the first quadrant contain raised circular perimeters to facilitate a determination whether an appropriate volume of whole blood has been deposited thereon.

26. The blood plasma separator device according to claim 23, wherein the first and second quadrants each include from 2-4 spots arranged in one or more rows.

27. The blood/plasma separator device according to claim 23, wherein a length of the first and second quadrants constituting the elongated bifacial sheet according to the first configuration is from about 50 mm to about 250 mm, and a width of the elongated bifacial sheet constituted by the quadrants is from about 20 mm to about 60 mm.

28. The blood/plasma separator device according to claim 23, further comprising a separate and independently foldable quadrant for deposition and storage of additional whole blood or plasma specimens for further analysis.

29. The blood/plasma separator device according to claim 23, wherein the device is configured as a hand-held point-of-care device and is compatible for use with smart phones.

30. The blood/plasma separator device according to claim 29, further comprising identification technology on an outer surface thereof when in a fully folded/closed configuration.

31. The blood/plasma separator device according to claim 30, wherein the identification technology provides for HIPAA-compliant cloud-based transmission and storage of personal identifying information of a patient associated with the biological specimens collected and stored in device.

32. The blood/plasma separator device according to claim 31, wherein the identification technology comprises an authentication feature that enables authentication of specimens stored in the device.