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    • 专利摘要:
    DEVICES AND METHODS FOR BLOOD PLASMA FILTERING. The present invention provides systems, devices, kits and methods for separating blood plasma from whole blood. Specifically, the present invention provides systems, devices and methods for separating a fixed volume of blood plasma from whole blood with minimal energy consumption.
    公开号:BR112013002064B1
    申请号:R112013002064-4
    申请日:2011-07-27
    公开日:2020-11-24
    发明作者:Arman Nabatiyan；Ashley Marie Yanchak Boggiano；Samuel John Pickerill；Sujit Jangam；Shivani Gupta；David M. Kelso；Kunal Sur
    申请人:Northwestern University；
    IPC主号:
    专利说明:

    This application relates to the priority of the Hedi of the US Patent No. Seris 61 / 368.1 *) 6, filed on July 27, 2'313, which in this matter is 1 in the orpora of the reference by its totality. FIELD OF THE INVENTION
    The present invention provides systems, devices, kits and methods for separating blood plasma from whole blood. Especially, the present invention provides systems, devices and methods for separating a volume (e.g., fixed volume) of blood plasma from whole blood with minimal energy consumption. FUNDAMENTS
    Several upstream processes are necessary before. a complex biological fluid can be analyzed for analytes. For example, to perform HIV viral load detection, separate between plasma and blood is the first step upstream since homoglobin and blood cells are interfered with amplification and detection of viral RNA. Plasma separation is also a decisive process upstream for the detection and diagnosis of infectious diseases. For example, detecting HIV in adults using specific anti-HIV antibodies or detecting I1T.V in liquids using a p2 / i nucleic acid and HIV requires separation of blood plasma. total.
    Under laboratory conditions, the separation of plasma from the Lol.al blood is carried out by centrifuging the blood by z.Q. In this way, the solid blood components are located in the sediment and the supernatant liquid consists of plasma. This protocol usually requires a trained technician to manually extract the supernatant by pipette for further analysis. Although large-scale automated sample propagation systems can eliminate the manual step, these devices are expensive instruments, making them unsuitable for testing with limited resources or portable t and s t e equipment.
    All ten side developed to integrate blood separation by centrifuge with other steps downstream using a microfluidic platform (Brenner et al. Special Publication-Royal Society of Chemistry 200'4, 296: 566-568; Haeberle et al Lab on a Chip 200 6, 6: 776-781; Kang et al. Special Publication-Royal Society of Chemistry 2004, 2 96: 614-61 6; M.adou et al. Biomedical Microdevices 2001, 3: 245-254 ; Toner & Irimia. Annual Review of Biomedical Engineering 2005, 7: 77-103; Luo et al. J Clin: Microbio / 2005, 4 3: 1851-1857, in this subject incorporated by reference in their totality). However, these methods work with an extremely limited volume of blood, require the use of an instrument to create the forgacentrifuge, tend to clog by waste and / or provide only limited purity. The use of synthetic membranes to separate blood from plasma avoids some of the problems caused by centrifugation and microtiter systems; however, the devices are complex in each other. the reason for the need for multiple filter layers (Vogel el. al. Boehringer Mannheim GmbH, 1.984; Vogel et al. Boehringer Mannheim GmbH, 1989, incorporated in this regard by reference in their totality), inherently sacs and the membrane materials employed are difficult to use (eg, fiberglass), comprise materials that interfere with analysis (Baumgardner & Loewen. Ahlstrom Filtration, Inc., 1993; Loewen & Baumgardner. Tappi Journal 1996, 79: 183-184, in this subject incorporated by reference in their entirety) and contains materials that slow the flow of blood to the filters (Suzuki & Ho: A magnetic force driven chaotic micro-mixer, PP. 40-43, 2.002: 40-43, in this subject incorporated by reference in full water). SUMMARY OF THE INVENTION
    In some embodiments, the present invention provides a blood plasma filtration method comprising (a) providing: (i) a filter module, wherein the filter module comprises a filter configured to allow the passage of blood plasma but not others blood components; (ii) a plasma collection module, in which the plasma collection module comprises a collection membrane configured to extract blood plasma to the plasma collection module by capillary action; and (ii: i J a blood sample; (b) administering the blood sample to the filter of the filter module; (c) placing the collection membrane of the plasma collection module in direct contact with the filter of the filter module; and (d) extraction of blood plasma through: from the fill.rg.to the plasma collection module. In some modalities, the feed is passive. In some modalities, the extraction does not require any electrophoresis. , centrifugation or Lensao greater than the pressure aLmosfer.i ca. In some modalities, the filter module accommodates a fixed volume of the blood sample In some modalities, the plasma collection module accommodates a fixed volume of plasma. , the fixed volume of the m / xiulude Plasma collection is less than the fixed volume of the filter module In some embodiments, the fixed volume of the plasma collection module is independent of the blood sample volume. of the plasma collection module does not depend on the hematocrit of the sample blood. In some embodiments, the filter comprises a VIVID GF membrane. In some embodiments, the collection membrane comprises an AHLSTROM 142 fiber collection pad. In some embodiments, the collection membranes comprise a PALL A / D fiber collection pad. In some embodiments, the method further comprises (e) a plasma supply in the plasma collection module.
    In some embodiments, the present invention provides a device for separating plasma from whole blood comprising: (a) a filter module, wherein the filter module comprises a filter configured to allow passage of blood plasma but not other components blood; and (b) a plasma collection module, where the plasma collection module comprises a collection membrane configured to extract blood plasma into the plasma collection module by capillary action. In some embodiments, the filter module accommodates a fixed volume of whole blood. In some embodiments, the plasma collection module accommodates a fixed volume of plasma. In some embodiments, the fixed volume of the plasma collection module is less than the fixed volume of the rilt.ro module. In some ways, the filter comprises a VIVID GF or VIVID GR membrane. In some embodiments, the collection membrane comprises one. fibru collection pad AHLSTROM 142, l-’Al.l. ACCUWIK or PALL A / D. In some embodiments, the device comprises PMPs in the collection module. In some embodiments, the device comprises analysis reagents in the collection module. Additional features are described here. DESCRIPTION OF THE FIGURES
    Figure 1 shows a lateral view that expands the blood attachment point on the membrane, the blood side and the plasma side on the membrane.
    Figure 2 shows a configuration of the side flow showing. the sample cushion that separated the blood cells from the plasma, the collection cushion that collects a fixed volume of plasma and an absorption cushion to collect the extra plasma.
    Figure 3 shows a vertical flow arrangement; the cell separation and plasma collection membranes are clamped between an upper plastics blade and a base blade.
    Figure 4 shows the chemical structure of the Merquat poll cation.
    Figure 5 shows a graph of cyclical threshold (CT) versus log1-0 (copies of HTV-1 viral RNA): the squares denote the C values, obtained when a plasma sample containing 11.1V—] was purified using the IFF method; the lozenges denote the CT values obtained when V'Fl Cod is used to collect whole blood plasma containing HIV-1 and Ahlstrom 142 fol used for plasma collection.
    Figure 6 shows a cross-sectional view of columnar fragments made by 1 OX technologies showing pillars that are 14 pm high and stick size from 152 pm.-
    Figure 7 shows an illustrative drawing of the Bangui neo plasma unit. The sanguine® plasma unit has two parts, a ceil Las Vegas module and a plasma module. After separation, the plasma module is removed and has been transferred to the cartridge for processing in the system analyzer.
    Figure 8 shows an illustrative first generation plasma separation and collection module: (left) PALL VIVID GR cell separation membrane connected with the separation module; (in the center) plasma collection membrane PALL ACCUWLK connected with the collection module; (on the right) separation and collection modules connected to each other to separate plasma from whole blood.
    Figure 9 shows second generation plasma separation and collection modules: (from left to right) the cell separation module- connected to the PALL VIVID membrane with the adhesive, the anti-rotation accessory connected to PALL ACCUWLK and the device threaded positioning.
    Figure 10 shows a positioning mechanism for studies of the blood-plasma unit pressure.
    Figure 11 shows a graph.Lco of the cyclical threshold of the volume of plasma collected. (squares) and nsLrrt for 600 Cop Las of HIV-1 virus per ml of blood (tri-angles) VOZSJUS volume of whole blood added to the sanctioning unit Lasma.
    Figure 12 shows a graph of the volume of plasma collected on the PALL ACCDWIK membrane (plasma collection module) by; by adding 125 μl of blood to the blood-plasma unit in t. different sizes of the VIV I D.
    Figure 13 shows a graph of the cyclic ll.mJ.ar of the volume of plasma collected (quad fades) d ostimized for 600 plots of HIV-1 virus per ml of blood (cy reales) versus blood hematocrit levels (%).
    Figure 14 shows a graph of the separated plasma volume (uL) of 125 pL of whole blood using the plasma separation and collection module versus the blood hematocrit (percentage). The squares denote the average amount of plasma obtained from a different blood sample measured in duplicates. The Sol Ida line denotes the average volume of blood collected from all samples and the dashed lines show the mean ± 2 standard deviations.
    Figure 15 shows a schematic drawing of the IPF processes: (left) 1 PM P's bind to NA and are removed by a magnet outside the Use cap for the liquid wax elution cap .; (the right) molded cartridge containing Lysis cap, e cap) and a colored liquid wax.
    Figure 16 shows a graph of cyclic Jimiar (CT) versus log10 (copies of HIV-1 per pL of plasma). The squares denote separate samples using the separation module and the plasma pool and the. oval shapes denote separate poles used by the center.
    Figure 17 shows more R'T-PCR quanti LaL i va for HIV-1 plasma with Ambion PMPs stored in the membrane. ACCUWIK: the standard curve of the pure 4 d valves Lterenl.es RNA concentrations represented graphically versus the log10 of the number of HIV-1 viral capillaries. The solid squares are the CT values.
    Figure 18 shows a graph of Bl and-Altmar, comparing the samples with BMPs in Lysis Lampsp and in the ACCUW.IK membrane: quadratics know how to differentiate between the method dots.
    Figure 19 shows that the blood separation and plasma collection membranes do not significantly attract p24 protein analyte.
    Figure 20 shows that - no specific analyte signal loss is observed in plasma collected from whole blood with separate analyte samples with a device of the present invention.
    Figure 21 shows that no loss of specific analytical slna.l is observed in plasma collected from whole blood with analyte pices separated with a device of the present invention.
    Figure 22 shows that the signal production :.) of the desiccated collection cushions with analytical specific detergents is comparable to reactions with dissolving detergents.
    Figure 23 shows that the signal production of desiccated collection cushions with ana Li se's specific detergents and protein analyte is comparable. to reactions with detergents and analyte in solution.
    Figure 24 shows a left-hand drawing of a plastic component peeling off the vol (from uni.co) in the form of a shell, part of the filter element. The Integra shell has its own soparaqap filter and inLorfuce marker with a GlcmenlQ extdn of the plasma collection, which now runs the collection tube.
    Figure 25 shows: (a) the upper half of a shell is folded and closed. The right and left locking algae hold and keep the shell closed, as shown in the cross-sectional projection in (b).
    Figure 26 shows (on the left) a schematic drawing of the basin part of the shell. A separation filter is aligned and joined to the shell, here, by means of an energy manager using ultrasonic welding ;. and (on the right) a schematic drawing of the integrated shell and the separation filter. The energy management plastics merge with the separation filter and surround it, creating a pseudo-ring barrier.
    Figure 27 shows (on the left) a schematic drawing of a closed and fastened shell, which has a separation filter connected to it (eg, by means of ultrasonic welding and using a power manager) ; and [the right) is a cross-sectional projection of the schematic drawing on the left showing that the separation filter is connected so that when the shell 6 is closed, it is flush with the lower half of the shell.
    Figure 28 shows (a), in the schematic drawing of the plasma collection element in the top view, a pias.tica tape for testing the substrate with a small portion of bilateral adhesive comparing with diagnosis. i co, quo Connects to the ribbon collection mombrana for the taste of substrate; (b) side view of the collection element; (c) projection of the enlarged lateral wist (angle in lines 1. activated) showing the fold, the perforation and the double layer of adhesive that so.11 g the tape for Lqstc of substrate o Si membrane of collect.
    Figure 29 shows (S left) schematic drawing of the plasma collection element and the filter element with its respective membrane and filter; and (on the right) the collection element fitted to the filter element by the phenotype and aligned with the rear wall.
    Figure 30 shows (on the left) the collection element integrated with the filter element, where the upper half of the shell was lowered and locked by the locking algae. (on the right) this allows the separation filter and collection membrane to overlap and compress, which facilitates the separation of blood.
    Figure 31 shows (on the left) a schematic drawing showing the assembled module for blood separation; and (right) a transparent side view of the assembled module showing the flap that fits with the fold in the line for testing the substrate of the collection element.
    Figure 32 shows an overview of the stages of the pre-pressure of the blood separation using a filter element and a plasma collection element of the shell type: (a) distribution of the sample if blood in the separation filter; (b) the separation filter leads the plasma to the collection membrane; (c-d) the operator compresses the lower half of the left and right locking algae to open the upper half of the filter element; e. (e) the filter element and the collection element are separated from each other and can be used for subsequent testing processes (eg, CD4 count, PCR, immunoassay) - DF.FINIQOES
    For fuel to understand the understanding of the invention, in several Lermos and expressions defined below:
    As used herein, the term "individual" refers to organisms to be treated by the methods of the present invention. Such organisms preferably include, but are not limited to, mammals (e.g., murines, simians, horses, cattle, suites, cards, felines, primates etc.) and mats preferably to humans. In the context of Invenpao, the term. "individual" typically refers to the person providing the sample or from whom the sample was taken (eg, tissue sample, fluid sample [such as blood sample]). The term "patient" is generally used when referring to a human individual.
    The term "diagnosed", as used herein, refers to the detection of a disease by six signs and symptoms (e.g., resistance to conventional therapies) or by genetic analysis, pathological analysis, histology analysis and the equivalent.
    As used herein, the term "in vitro" refers to an artificial environment, and to processes or reactions that occur. within an artificial environment. In vll.ro environments include, but are not limited to, test tubes. The term "in vivo" refers to the natural environment (e.g., an animal or a cell) and to processes or reactions that occur within a natural environment.
    As used here, the term "sample" is used in its mats ample sense. A sample can comprise a cell, tissue or fluids (eg, blood, plasma, etc.), ions, cell material, qendmic DNA (in solution or attached to a solid support, for example, Southern analysis biot), RNA (in solution or attached to a solid support as an example of Northern blot analysis), cDNA, (in solution or attached to a solid support) and the equivalent®.
    As used here, the terms "iso1ar" or "iso1ado" refer to:: only to a cue and identified sample separated from at least one component or contaminant® to which it is ordinarily associated in its strength. Natural Lu. An isolated sample is present in a form or condition that is different from that in which it is found in nature. In contrast, a non-isolated sample is found in the state in which it exists in nature.
    As used herein, the terms "purified" or "purifted" refer to the removal of components (e.g., contaminants, unwanted components) from a sample. As used herein, the term "substantially purified" refers to molecules that are at least 60% free, preferably 75% free and more preferably 90%, 95%, 99%, or more, free of other components in the quads it it is usually associated. DETAILED DESCRIPTION OF THE INVENTION
    The present invention provides alternative devices, kits, methods and methods for separating blood plasma from whole blood. Specifically, the present invention provides systems, devices and methods for separating a volume (e.g., a fixed volume). of blood plasma from sa.ngue .total with mini energy consumption. In some embodiments, the present invention turns systems and devices for draining blood from whole blood plasma. In some embodiments, devices separate. blood plasma from other blood components (eg blood cells). I: In some modalities, devices purify blood plasma. In some embodiments, devices isolate blood plasma. In some embodiments, devices concentrate blood plasma. In some embodiments, devices effectively separate and concentrate a fixed volume of plasma from whole blood. In some embodiments, the invention separates, isolates, putifiates and / or concentrates blood plasma from whole blood for additional energy consumption (e.g. ., without heating, cent.rifugag.ao, electricity etc.). In some embodiments, the present invention provides a filter element and a collection element. In some embodiments, whole blood (eg, not fl 1. trade) is added to a filter element, and blood is aspirated (eg, by capillary action, gravity, etc.) through the element filter element for the collection element. In some embodiments, the present invention provides a filter element. Some modalities err, one or more blood components (eg, cellular c-omponen les) move slowly through the filter element of the blood plasma. In some embodiments, blood components from the plasma (e.g., cellular components) move more slowly through the filter element than plasma. For some time, one or more blood components (eg, cellular components) are unable to move through the fi rrp element. In some embodiments, blood components are excluded from the plasma (e.g. cellular components) are unable to move through the filter element. In some embodiments, blood plasma rapidly Cp. faster than other blood components.) advances through the filter element towards the filter element and / or the element. filter. In some embodiments, the filter element comprises a filter, a membrane, a matrix and / or a pad capable of separating blood plasma from other blood components based on capital. Although the present invention is not limited to any specific mechanism of action and an understanding of the mechanism of action is not necessary to practice the present invention, the movement of a liquid through a material by capillary action is controlled by the equation:
    where y and the superficial lensSp at the liquid-air interface (energla / ark), 0 and the contact angle, by the density of the liquid (mass / volume), ge acceptance due to gravity (length / time'- ') and area trajer.dria of the drain through the material (length). Therefore, different liquids move through a material at different rates based on the surface tension at the liquid-air interface and the density of the liquids. In some embodiments, plasma moves more rapidly through a material (eg, filter element) than other blood components (eg, cellular components). In some embodiments, the present invention separates blood plasma from four components of the total blood based on differences in the rate of gap movement through a filter material. In some embodiments, a filter element comprises a filter and / or membrane secured to allow the passage of blood plasma and other components of whole blood at different rates. In some embodiments, a filter element comprises a tie-plasma separation membrane. In some embodiments, a filter element comprises a VIVID Plasma Separation Membrane. In some embodiments, a filter element comprises a VIVID GF Plasma Separation Membrane. In some embodiments, a filter element is configured to separate a defined volume of whole blood (eg, 15 pL, 25 pL, 50 pL, 75 pL, 100 pL, 125 pL, 150 pL, 200 pL, 300 pL, 400 pL, 500 pL, 750 pL, 1 ml, 2 ml, 3 ml etc.,). In some embodiments, the filter element comprises a membrane of any suitable size. In some embodiments, the filter element comprises a membrane of any suitable diameter (eg, 5 mm ... 8 mm ... 10 mm ... 12 mm ... 15 mm ... 18 mm. .. 20 mm... 25 mm.. 30 mm... 40 mm ... 50 mm ... etc.). The present invention is not limited to the materials used in the filter element, and any material inferred by a person in the field who provides suitable filtration qualities can find use in the present invention. In some embodiments, a filter element is disposed of. In some embodiments, a filter element is reusable. In some embodiments, a filter comprises a molded shell (eg, disposable molded shell) (SEE FIGS. 24-25). In some embodiments, a fill element, rq, comprises a shell (eg, single-use shell) and a seperation filter (VER FISH. 25-27). In some embodiments, a component of the shell and mol is given by injection (from a molded plasl.J.ca resin). In modal modalities [dudes, a filter element. uomproonde a shell component integrated with a s & parapao filter. Presentations, invention is not Jimi La to the materials or the configuration used in the filter element, and any material inferred by a person in the field - which provides adequate filtration quality may find use in the present invention.
    In some embodiments, the present invention provides a collection element. In some embodiments, a collection element comprises a substrate, a pad, a matrix material and / or a filter. In some embodiments, a collection element is configured to collect a fixed plasma volume from a whole blood sample: (eg 10 pL, 20pL, 30 pl .., 40 pL, 50 pL, 60 pL, 60 pt, 30 pL, 80 pL, 90 pL, 100 pL, 200 pL, 300 pL, 400 pL, 500 pL, 1 ml etc.). In some embodiments, a collection element comprises a plasma separation membrane, a collection pad, a collection matrix, etc. In some embodiments, a collection element comprises one or more materials such as fiberglass, polyester, nitrocellulose and / or cellulose. In some embodiments, a collection element comprises one or more materials selected from the WHATMAN Fusion 5 membranes (Whatman Diagnostic Catalog 2010, page 8; in this subject already incorporated by reference in its entirety), PALL ACCUWIK, PALL A / D, AHLSTROM 111 (Ahlstrom Laboratory Products Catalog (2009), page 15; in this subject in body by reference in full age), AHLSTROM 151 (Ahlstrom. Laboratory Products Catalog (2009), page lb; in this subject. Incorporated by reference in its entirety) with or AHLSTROM 142 (Ahlstrom Laboratory Product is Catalog (2009), page 15; in this matter incorporated by retention in its entirety) with / or any other suitable membrane, cushions or matrix materials known. for aquoles with practical experience.Lea. In some embodiments, a collection element © comprises a membrane: collection AHLSTROM 142. In some embodiments, the filter element comprises a membrane of any suitable shape (eg 2 mm ... 4 mm ... 6 mm ... 8 mm ... 10 mm ... 12 mm ... 15 mm ... 20 mm ... 30 mm ... 40 mm ... 50 mm. ... etc. ...). In some embodiments, a collection element comprises a substrate test tape, a double-sided diagnostic adhesive and a collection membrane. In some embodiments, a collection element comprises a collection membrane. In some embodiments, a collection element comprises a double-sided magnetic adhesive. In some embodiments, a collection element comprises a substrate test tape. In some embodiments, a collection element comprises a substrate test tape, a double-sided diagnostic adhesive and a collection membrane. The present invention is not only limited to the materials used in the collection element or its configuration, any material or configuration specified by a person in the field as it provides suitable collection qualities may find use in the present invention. a filter element and a collection element comprise an imca unit. In some modalities, a filter element and a collection element comprise separate collection and filter modules. In some embodiments, a separate module and a filter module are optional connected and / or connected. For some modalities, an IJJt.ro module comprises a filter element. In some embodiments, a filter element (eg membrane) in a filter insert is replaceable and the filter module is reusable with a new filter element (eg membrane). In some embodiments, a collection module comprises a collection element. In some modal I data, a collection element (eg membrane) with a collection module is replaceable, and the collection module 6 is routable with a new collection element (eg membrane).
    In some embodiments, the present invention provides a matrix within a collection module for collecting blood (e.g., from a digital needle, from a vein, etc.). In some embodiments, the present invention provides a matrix to elicit blood by digital puncture. In some embodiments, the present invention separates plasma cells when whole blood is added. In some embodiments, the amount of blood needed is minimized by maximizing the efficiency of plasma separation.
    In some embodiments, devices of the present invention provide for separation of components from whole blood (e.g., blood cells) from plasma. In some embodiments, the plasma and separate cellular components can each be used for sub-sequential applications (eg, tests). For example, blood cell components are separated from the plasma to monitor CD4 counts, and the plasma component is used to determine viral load. Using both cellular and cellular components. plasma, devices of the present invention provide samples for multi plus different analyzes (VI -. 'R PIC. 7) .i'-have some modalities, systems, methods available to provide different components between components of plasma and cellular components in the sanguetotal. In some modes, plasma separation from other blood components is completed in less than 1 hour (eg, 45 minutes ... 30 minutes ... 20 minutes ... 15 minutes ... 10 minutes ... 8 minutes ... 6 minutes ... 5 minutes ... 4 minutes ... 3 minutes ... 2 minutes ... 1 minute ... 30 seconds).
    In some modalities, systems, devices: emetodos and raise a fixed amount of plasma. In some embodiments, systems, devices and methods carry a fixed amount of plasma on a membrane, a filter and / or pm cushion (eg 71HLSTROM 142). In some embodiments, the present invention collects a fixed volume of plasma and therefore provides a modification device. In some embodiments, devices of the present invention carry a fixed volume of plasma, regardless of the volume of blood added. In some embodiments, devices of the presence of the invention carry a fixed volume of plasma, independent of the hematocrit of the added blood. In some embodiments, systems, devices and methods of the present invention carry a fixed volume of plasma from a whole blood sample (eg 10 pl ,, 20 pL, 30 pL, 40 pL, 50 pl ,, 60 pL, 7G Ilk W SB pin 3 00 ut, 200 gt., 3.00 pL, 400 pl ,, 500 pL, 1 m, I. etc.). There are some modalities, systems, devices and methods of the presbytery that are approximately 50 liters of plasma, blood. In some embodiments, devices of the present invention provide collection of a volume of plasma seionated to provide a desired viral load in the sample (eg, a volume of 50 pl. Provides a sensitivity of 600 copies of IVIV / ml of blood for detection of viral load by RT-PCR).
    In some embodiments, devices, systems and methods of the present invention operate pass! again., requiring no active pumping, gentrifugation, electrophoresis process. and / or eietricity to operate. Rm some medals through capillary action (modalities, devices for the provision of laboratory, field) and / or borderline dose-passive and / or gravity. dolocalizegoesleito.plasma emlocal! zagaoremotas (P-no
    In some embodiments, a membrane collection element.) And mcr.tr which: p membrane separation element). In some embodiments, a device and / or a system in which the collection element (eg, membrane) is smaller than the separating element (eg, membrane) serves to concentrate plasma and has a small volume. In some embodiments, the concentration of blood plasma in a collection element is useful for anaerobic steps if the laboratory requires increased sensitivity (eg, an immunoassay or for improved capture of nucleic acid in a paramagnetic particle).
    In some embodiments, a device of the present invention, a collection eJemenlu and / or. a moduilio of. colc'ta Supplyri and / o'u serve as a blood plasma storage device. In many ways, the presence of the invention provides a storage element for blood plasma. In some embodiments, blood plasma is stored in the collection element (p. © X., Collection matrix). In some embodiments, plasma is transported while contained within the collection element. In some modifications, a filter element containing filtered blood components is discarded prior to storage and / or transport'd of blood plasma.
    In some embodiments, a device of the present invention, a collection element and / or a collection module are integrated with downstream processes (eg, caxtucbo or other device for extracting, purifying and amplifying HIV viral RNA; a lateral flow system for the detection of anti-HIV antibodies or p24 protein; etc.). In fashion 11, some device or system of the present invention comprises modules downstream for additional analysis or manipulation of separate blood plasma. In some fashion, ages, a collection element allows extraction of plasma collected directly into the cap gas for further analysis and / or handling. For viral load procedures, the plasma collection membrane allows extraction of the viral RNA present in the plasma directly into the cap (eg, ii.se/ cap) Lgapgo Ambion [Applied Biosystem,]) for further processing downstream. In some modalities, the direct extraction of the collection element eliminates manipulation steps associated with other protocols that may result in loss, contamination or dariQ of the sample. For side-flow procedures, the plasma collection membrane allows extraction of the stem cells (eg, anti-HIV esped ari-ti-corpus 1 fi Cos, core p24 protocols, etc.) for further downstream processing ' . In some embodiments, analysis reagents may be stored (eg, dried or 1 liter) in the plasma collection module. In some embodiments, a collection element provides a matrix for storing (eg, long-term storage) parti.eu.las paramaqn6ticas (PMPs). In some embodiments, PMPs are used to capture nucleic acids (eg, viral RNA) and subsequent processing. In some embodiments, a collection element allows PMPs to be extruded out of the membrane using a magnetic force (eg, generated by a permanent magnet or an electromagnet), eliminating the need for agitation and centrifugation of the solution, steps usually associated with samples collected on filter membranes for PCR procedures.
    In some embodiments, kits are provided that comprise one or more or all of the necessary, sufficient or useful components for preparing or using any of the devices described herein. In some embodiments, the kits comprise control reagents, instructions (eg, software), data analysis devices or any other desired components. Example 1 Separation method
    Lateral flow separation. Three different side-flow membranes, namely, Pall Cytosep 1662, Watman Fusion 5, Whatman MFI, have been tested for the separation of plasma in a lateral flow (see FIG. 1) The membranes were connected based on the manufacturer's recommendations, on the capacity of the plasma for 100-200 pL of whole blood without obstruction, pore size and filtration capacity. of the membrane, the plasma side moves more quickly than the blood side (see Fl Cl 1). The blood section can be terminated by cutting the red membrane sequence. The volume of plasma collected was determined by weighing the cape .. of filter before and collection depots p res s upondo-s and be the plasma density of 1025 kg / mJ (Benson K: MACAT review.Emory University 1999: 1.41-1.99, in this incorporated seat by reference in their totality.) Using a blood sample collected by venipuncture with a 50% hematocrit, the elevated plasma ions was observed with. the Cytosep membrane by 75%. Although this percentage of recovery can be improved by adding a collection pad with a higher cap & nuftier (households (SEE FIG.2), this method required the following limitations: (1) the flow rate it was slow, making the process time consuming, ( ) apposition on the side of the blood varied according to the hematocrit, the volume of blood added and other re-genic properties of the blood, requiring the user to manually locate the blood donor and section the membrane to separate plasma from whole blood, (3) the collection of a fixed volume of plasma would also require deviation out of a fixed length of membrane containing plasma, which depends on the capacity of the membrane. The need for the deviation step was eliminated by employment of an alternative configuration) (SEE FIG. 2), .when blood and co ", elude in a sample cushion that prevents Banguinea cells from flowing into the collection cushion. The collection cushion has the appropriate size opposite - pope eject a pl a.sma extra r'fu For the cushion modalities of the present invention demonstrate that the 24/69 side of the red blood cells does not stop completely, and none of the aforementioned membranes prevents complete extravasation of the cells to the collection cushion. To eliminate this problem, the sample cushion was sized enough to accommodate the variance of the blood volume associated with the digital puncture collection. However, this reduces the efficiency of plasma collection. Another solution to the problem was to add an exact amount of blood to the sample pad. This can be achieved by first collecting the blood in a capillary tube with an indicator for blood volume and then attaching it to the sample cushion. However, this method adds an extra step to the process. Separating the vertical flow tie. Plasma separation was performed in a vertical flow configuration (SEE Fid 3), hematiae and leukocytes are captured by the separation membrane thanks to their small pore size while the blood plasma components flow through the collection membrane. . The plasma flow rate, Q, directly provides1 the cross-sectional area of the flow and inversamerite proportional to the flow length. Due to the large cross-sectional area and the small length of the flow in comparison. lateral flow, the separation between plasma and cells is faster in vertical flow than in lateral flow. Example 2 Membrane
    Membranes were located based on strict criteria, ie, exl. plasma d; whole blood30 and RNA expression capacity of the membrane in the Lysis / binding cap (Applied Biosystem, Carlsbad, GA) containing ethanol and guanidinium thiocyanate (GuSCN). Experiments were carried out during the development of modalities of the present invention to determine the efficiency of plasma separation from whole blood and sample collection for specific membranes (See FIG. 3). FALL Cytosep 1662, Whatman VF1 and PALL VIVID GR were tested for plasma separation and Ahlstrom 111, Ahlstrom 142, WhatmanMFI and PALL ACCUWIK Ultra were tested for the sample companion.
    Separation from plasma. The separation membrane was sized for 100 pL of blood ■ following the manufacturer's recommendations. The collection membrane was dimensioned to 50 pL of plasma using the membrane capacity recommended by the manufacturer. A fresh blood sample containing anticoagulant was used. The blood sample hematocrit was adjusted to 40 ° by aliquoting a known amount of plasma and cells in a microcentrifuge tube. The plasma and cells were mixed well in a mixer for 20 minutes. The filter membranes were weighed and then configured for testing (SEE FIG, 3). Pressure was applied by adjusting the screws to promote contact between the membranes. 100 pL of blood was added to the soup membrane, left to stand for 10 minutes. The collection pad is subsoquontemenl.e plated again. The volume of plasma collected was calculated by establishing the difference between the pre- and pos- sible weights d.Lvidida by the density of the plasma. Plasma density was estimated at 102.5 kq / m3 (Benson K review: MCAT. Emory University 1999: 141-199, in this subject incorporated by reference in its entirety), which is an approximate average value. The plasma recovery was then calculated by dividing the volume of plasma collected by the total amount of plasma: na. original sample. The separation and collection membranes were placed in a biologically hazardous container. The set is washed with 70% ethanol, dried and then used for further study.
    The results of experiments carried out during the development of modalities of the present invention suggest that VFI and VIVID GR demonstrated ideal properties for plasma separation and 111, 142 and ACCUWIK Ultra exhibit useful properties in plasma collection (SEE TABLE 1).
    TABKT.A 1: PERCENTAGE OF PLASMA RECOVERY OBTAINED WITH DTFERENT COMBI NATIONS OF MEMBRANES OF SF.PARACAO AND COLLECTION. Vlfil 1- VFI REFER- CK TO USING L / W! VF1 PiXADAS BETWEEN SIL IN "ARTE SURER I OR, Vi-'l WITH MIWUAT REFKRE-.3E A VHi CCWTHNlio JAT .. VI-1 I G931-AII RKFERE-BE- AS- DIMS MEMBRANAS- FIXADASJ KNTKF S '; W PART SUWRlaR.
    The separation factor is controlled by several factors. The number of capillaries DU or Larnunbo in the pore of the collection membrane interferes with the extradition. The smaller the size of the pore, the greater the extraction efficiency, which would indicate greater efficiency by using the '111 membrane compared to the 14-2. Although the separation membrane must separate plasma without obstruction by geT formation or clogging, its capacity must be minimal. A membrane with high liquid capacity would result in S lesser extraction efficiency in the collection membrane.
    The VF1 membrane consists of pores of varying sizes and only retards the flow of the membrane. Considering the time span sufficiently long (> TO minutes), consequently the cells will leak from the membrane to the collection pad. The VF1 + G934mAH membrane is a combination of a separation pad and a clay membrane and only works with collection membranes with small pores such as 111. The barrel membrane has small pores of 3 pm and prevents passage of the cells. VF’l is important to prevent the clay membrane from becoming clogged. The use of MFI as a collection membrane slows down the flow sufficiently to prevent any swelling from moving LentameriLe to the collection membrane, however, it also reduces extraction efficiency.20 How. Alternatively, the VF'l membrane was immersed in a 2% well solution. 1 ion ion, Merquat 550 (Nalco Company, IL), (VhiR FIG. 4).
    ELluipao. Were carried out and, during the development of fashion.1 ages d, the presence of the Invention to determine the most suitable membrane for RNA extraction from the sample collection membrane in the cap 1 i se./Li gagao . Can the effi ciency of the classical efficiency of nucleic acid be be affected by the Link. of the nucleic acid with the RNA in the presence of the l.i se / binding cap containing high. ragAe concentration of GuSCN 30 and isopropanol. Membranes with extremely small pore sizes may allow viruses to diffuse into the membrane from the separating membrane, but they may not allow the viral RNA of the virus that has undergone lysis to diffuse out of La when the membrane is immersed on the cover of li.se/liqacao.
    The separation membrane was sized to 50 µl of the plasma using the manufacturer's recommendations. The HIV-1 virus, purchased from Rush Virology Quality Assurance Laboratory in the ratio of 1.5: X 106 cop.Las / ml of plasma, was diluted in seronegative plasma to obtain HIV-1 concentrations of 300 copies / pL. The plasma sample containing HIV was added to the filter membrane and left to rest for 5 minutes. The filter membrane was placed in a microcentrifuge tube and 600 µl of lysis buffer was added to it and vortexed for 10 minutes. As part of the liquid © and absorbed into the filter membrane, 400 μl of the ise cap was removed, and a mixture; of beads consisting of 5 pL of Amblon paramagneLic particles (PMPs) and 5 pL of BindingEnhancer (Applied Biosystera; Foster City, CA) was added to it. This solution was vortexed for 4 minutes to bind the RNA to the PMPs. Calculation by Immiscible Phase Filler (TPF) was performed. Quantification of HIV-1 viral load was performed using the Abbott RealTimo HiV-1 Amplification Reagent. Kit (Huang et al. 'Nuc Acids Res . () 07, 35: el01, in this matterin co root a co pgr1 ref c rd ncLa in its entirety) (Abbott Molecular, Des Pl.aines, |: L) in volumes of the reaction of 25 pl. with the addition of 0.2 mg / mL of bovine serum rich albumin (B8667; Sigma), 150 mM trehalose (1'9531; Sigma) and 0.27, of "i'woon 20 (28320; Pierce Thermo Fisher Scientific) and 5 pL as Amplification reactions were performed on Cepheid SmartCycler 11 '(Sunnyvale, CA.).
    All membranes retained part of the RNA (Table 2). The PALL. ACCUWIK retained the least amount of RNA and also showed the least variability in the amount of RNA capture. Fiberglass membranes and cellulose membranes bound to RNA in the lysis cap.
    TABLE 2: MFIDIA OF LLM1AR Ct Ci .ICO (C,) AND DRRVTO. STANDARD OF C OBTAINED BY ELECTIONS OF VIRAL RNA FROM DIFFERENT FILTER PAPERS IN THE AMBTON LISE CAP, WITH SUBSEQUENT PURXFACTION BY I PL 'AND KTACAO I'M A CHAIN OF POLYMERASE BY TRANSCRIPTASE REVEKSA. WITHOUT MEMBRANE REFERRED TO A SAMPLE IN WHICH THE PLASMA SAMPLE FOLL ADDED DIRECTLY TO THE AMBTQN LISE CAP.
    Experiments were carried out. during the trial i v i m e of prescribing the invention to stop the loss of RNA in Ahlstrom 142. Blood Lola! containing H.LV-1 was separated using the procedure described in Figure 3.Whatman VFI was used for cell separation.sc Ahlstrom 142 was used for plasma collection, The RNA was oxL.r'aAdo and amplified pot using the protocol. © described above. As a control, plasma containing H.LV-1 was also purified by the TPF protocol. A difference of 1.5 ap CT would be expected between the plasma sample and the sample captured in Ah 1 sLrom 142 based on the results shown r ..-: Table 2; however, extra mud loss in C. 0.5 was observed (SEE FIG. 4). The additional loss was caused by the loss of viral particles in Vfl. A similar study carried out with separate plasma using VF1 containing Merquat did not provide any result, indicating that the virus was captured by the positively charged Merquat or that the Merquat transported with the plasma inhibits PCR even after purification by IFF. Example 3 Sample Collection Swabs
    Were. Experiments were carried out during the development of the present invention to examine the use of COPAN sterile swab applicators (Copan Diagnostics Inc., Murrieta, CA) as a sample collection device. These swabs will use a technology called flocculation that transmits thousands of sprays of nylon fibers from a perpendicular angle to the tip of the applicator. With this procedure for the collection, the liquid specimen is absorbed between the adjacent fragments of the nan 1 on filaments by Cap! I air. 'The specimens are 1 u i of the swabs one vck placed in a liquid medium. An additional benefit, in addition to more extensive collection, and how the Flocked fwabs Jiberam specimens more. quickly. The swab model includes a uex.ru ru like 1 han be a velvet brush. Although the swabs used for testing have been similar: immersion rods, the technology can be used to create a flake-coated membrane surface,
    HIV-1 virus, acquired from Rush Virology Quality Assurance Laborslory in the proportion of 1.5 X 106 copies / mL of plasma, were dll. in seronegative plasma to obtain an HIV-1 comparison of 300 copies / pL. The plasma sample was separated into the swab, and the swab was dipped in a microcentrifuge tube containing 400 µL of Use Ambion buffer, a mixture of beads was added and the sample was vortexed gently for five minutes and left at rest for 2 additional minutes. The PMPs were removed using an extraction tool and placed in a solution containing 200 µL of Ainbion lysis buffer also removed from the microcentrifuge tube. Purification by Immiscible Phase Filter (IFF) was carried out. Quantification of the HIV-1 viral load was performed using the Abbott RealTime HIV-1 Amplification Reagen Kit (Abbot! Molecular, Des Plaines, II.) In reaction volumes of 25 pL with the addition of 0.2 mg / mL albumin bovine serica (Sigma), 150 mM trehalose (Sigma) and 0.2% Tween 20 (Pierce Thermo Fisher Scientific) and 5 pi as a model. Amplification reactions were performed on Cepheid SmartCycler II (Sunnyvale, CA).
    Swabs are hydrophobic in nature and do not absorb. easily. As a result of the wear of the flakes, it has a low absorption capacity. Hritretantd, Lam is more suitable for slimy samples. The swab also absorbs RNA, and a difference in the (.> Of 2.0 fol observed between the samples deposited in the swab and the positive control in which the sample was deposited in the lysis buffer. This indicates that three quarters of the RNA was absorbed in swabs. Washing the swab with water, ethanol or lysis buffer before the test did not change the results. BMPs also tend to aggregate when they come into contact with the swab. swab in the presence of the Lysis / Bonding cap containing alcohol, salt and detergents, blunts the presence of the invention. It is not limited to any specific agony mechanism and an understanding of the mechanism of action is not necessary for the practice of the present invention.
    HIV-1 viruses, purchased from Rush Virology Quality Assurance Laboratory at a rate of 1.5 X 106 cdps / ml of plasma, were spread on the swab. The swab was immersed in 300 µl of Ambion eluigid buffer (Applied Biosystem; Foster City, CA) and left for 1 minute. As much liquid as possible was removed from the swab, 800 pL of the Ambion smooth cap was added to the aforementioned cap ■ containing the plasma samples, 800 pL was used instead of 40: 0 pL since the sample volume and large (350 pL). A large volume of elution solution was used, so that the swab could be completely submerged in the plug. 20 µl of bead mix containing 10 µl of PMPs and 10 µl of binding enhancer was added. PMPs were removed with an extracan stylus and placed in 400 µl of smooth plug. EPF purging was carried out with 50 pL of the elution cap. Quantification of HIV-1 viral load was measured using the. Abbott RcalTime HIV-1 Amplification Reagent Kill (Abbott Molecuar, Des plainer, IL) in reaction volumes of 25 pH with the addition of 0.2 mg / ml. of bovine series albumin (Sigma), 150 mM treaiosis (Sigma) G 0.2% of Tween 20 (Pierce 'Thermo Fisher Scientific.) and 5 pL as model ©. Amplification reactions were performed in Cepheid SmartCycler II (Sunnyvale, SA; using Roche TtH. As a positive control, the plasma sample containing HTV-i If. 'Was added directly to 300 pL of Amnion elution buffer without being applied and: t; a swab. As controls' negative, plasma negative for HIV-L and applied to the swab. Controls were processed in the same way as samples.
    More than half of the viral RNA is lost when samples are collected in swabs (Table 3). This loss cannot be attributed to the loss of elution cap removed with the swab. Part of the RNA or viral particles is absorbed in the swab.

    The usefulness of only using columnar fragments to collect it has been explored. Columnar systems are symmetric and not quite valid. I age in the dust associated with filter heaves. They are used for pesticide use only. tamiriaQroosmicroprismaticas reflective for high-gloss traffic signals (10X Technology, Libertyville, IL). The technology can be modified to create low-cost columnar fragments (eg, SEE FIG. 6). Preliminary® shields will show that their capacity for area unit f: oi is low due to the large diameter of the base and the small height of the column, making them unsuitable for use in high volume as is the use here. However, columnar fragments could be useful in procedures that require 10 <25 pL of sample. Example 4 Model Variables
    Experiments were conducted during the development of the present invention modalities for the creation of a filter paper separation system that can effectively separate and collect plasma and elute RNA (See Examples 1 - 3). The PALL VIVID GR Plasma Separation Membrane and the PALL ACCUWIK Ultra Medium filter and collect 80% of the volume of plasma contained in the whole blood sample 20 and eJuem almost all the RNA collected for amp! IEicagao, with a difference in the inner CT at 0.5 in compels ration with the direct plasma sample. Experiments were carried out during the development of the modalities of the present invention for the creation of a unit for soparagAo and plasma collection (PSC) by limiting the size of the membranes and the contact pressure between the two membranes using the VIVID membranes GR and ACCUWIK Ultra.
    Tatriarifiu of the VIVID Membrane. l VIVID Plasma Separation membrane is an Lm & trica, h i'droEdbiCa c 30 po l issu L phonic membrane with low biomolecular bonding. The asymmetrical nature of the material allows the cellular components of the blood (red blood cells, leukocytes and platelets) to be retained in large pores while allowing plasma to flow through the smaller pores to the other side of the membrane. The pulp components are filtered out without lysis, completely removing their con tarn loan ties. of plasma amosLr-a. As cellular components are collected inside the membrane, it is important to dimension the role of VIVID so that plasma is not trapped by the clogged pores. The blood volume capacity of VIVID is defined as the amount of whole blood per square centimeter of medium that can be quickly and effectively separated with minimal hemolysis. The blood volume capacity is directly related to the volume elimination of the material and is defined as 40-50 µL per square centimeter for the GR material grade. Considering the appropriate collection material, VIVID GR is fixed with a <80% plasma recovery rate. In a typical patient with a 50% hematocrit, 125 uL of whole blood would be required to collect 50 gL of plasma. Therefore, to ensure that no coagulation or use occurs, the VIVID membrane must have 2.5 cent, square meters of the area, which is related to a circular cut of 18 mm in diameter.
    Membrane size ACCUWfK. ACCUWIK 6 is used to collect the fixed amount of plasma. The absorption capacity as determined by Pa'll Life Sciences is 38 pl, per square centimeter, which corrodes a circular cut with 12.9 mm in diameter for a plasma sample with 50 pL. This small size of the ACCUWIK in relation to the size of the separation membrane VIVID allows concentration of the plasma in a small volume. The porous ACCUWIK provides a matrix for easy liquid transport and for additional chemical analysis within the matrix.
    Experiments were done to further confirm the useful size of the ACCUWIK membrane. The absorbency capacity of ACCUWIK Ultra was determined in pL per square centimeter by adding a fixed volume of plasma and determining the resulting occupied area. ACCUWIKUltra Medium is a fibrous hydrophilic membrane composed of hydroxylated polyester and characterized by uniform storms and quick release. The ACCUWIK strips were sectioned using a commercially available paper cutter. The width of each lyre was measured and recorded. The length of each lyre was approximately 1 mm mm. 50 pL of plasma was added to the end of the lyres, allowing lateral absorption until the movement of the front edge was stopped. The length of the filled area was measured and recorded. 100 pL of plasma was added to the end of a second group of strips, allowing lateral absorption until the movement of the front edge was stopped. The length of the filled area was measured and measured. I5 to determine the absorption capacity, the volume of plasma (50 or 100 pl.) 6 divided by the total area, filled, which is calculated mu I t i p I i i cting the length filled by the width of the lyre. Absorption capacities in pl. per square meter for the two groups of test strips were estimated. Although the reported capacity of ACCUWIK Ultra Medium has a side of 3-8 pL L / mm2, the average value calculated from the experimental test suggests that the capacity is actually slightly higher in the proportion of 4.5 ± 2.2 juL L / mm2., Using this absorbance capacity for the membrane, a circular cut with 12 mm diameters would be necessary to maintain the required plasma volume of 50 µL. Example 5 Plasma Collection and Separation Module
    The plasma collection and separation unit comprises two parts: the cell separation module and the plasma module. The module of. cell separation houses the VIVID filter, while the plasma module contains the ACCUWIK membrane (SEE FIG. 7). Once assembled, the membrane is in direct contact with the VIVID filter, providing capillary force to absorb plasma from the membrane. The contact pressure is established by the positioning of the plasma module with respect to the cell separation module. The whole blood sample is added through the opening at the top of the device and placed directly on the VIVID membrane. After an adequate amount of time has elapsed, the plasma module is disconnected from the cell separation module and inserted into the test cartridge for further processing. The cell separation module containing the filtered material can then be set appropriately.
    The first polypropylene roi generation system (SEE FIG. 8), Polypropylene was selected because it is chemically inert. However, polypropylene does not exist. Unsuitable mechanical properties. The filter membrane 6 is connected to the modules by a double-sided adhesive. Although various adhesion methods may find use with the device of the present invention, such as ultrasonic welding, thermal sealing, laser welding, clamping I: the mechanic etc., the adhesive was selected for the first generation device because it allows the plastic module is reused. The adhesive exhibited excellent bonding strength to most surfaces including low surface energy plastics such as polypropylene. The 0 ring on the collection module forms a waterproof seal and keeps the two modules together thanks to a cold adaptation. Preliminary studies conducted during the development of the modalities of the present fnv.engSo demonstrated that it was difficult to precisely control the contact between the two. membranes in this procedure which resulted in efficient separation of the plasma from the variable. The contact pressure was therefore identified as a key parameter to be optimized for consistent and efficient plasma separation.
    Second generation modules were constructed of polypropylene to incorporate a threaded locking mechanism that allowed the exact positioning of the filter membranes to each other (SEE FIG. 9). In order to prevent shearing of the membranes as the plasma module 6 is screwed to the Lota co module, an anti-rigid accessory has been designed (see FIG. 9). The cell ulus separation module has a pin-shaped pin that protrudes inwardly. The anti-rotation accessory has an inelasticity in which the pin in the form of a cavil is only resting. isl.o prevents the ant accessory. i rotate gl re, but allow it to move 11 v r onto nt © up or down. The vortical movement is further achieved by rotation of the threaded positioning device shown in the figure. Each module of the Lula cd separation and the corresponding threaded positioning device are configured to allow the dots to be attached. When completely starved, the distance "d" shown in Figure 10 is 1 mm. At this point, the stamping devices stamped on the cell separation module and the corresponding screw module device are located on the same vertical axis. When paired mimers are screwed together, the distance "d" is maintained. Example 6 Pressure
    Experiments were conducted during the development of modalities of the present invention to determine the ideal positioning of the ACCUWIK membrane in relation to the VIVID membrane so that permanent contact is maintained and the pressed ideal is applied to the entire interface of the Ciltro paper. The ideal placement of the filter paper was defined as the position that generates the highest volume of plasma collected with the least variability between tests.
    Filter papers and adhesives were sectioned to the appropriate size using a laser pot cutting machine. The modu '. those from cell separation and plasma were washed in IftOII to IS1 and allowed to dry. A circular adhesive of J 2 mm was used to secure the cut of ACCUWIK from 12. mm to the anti-fouling material. The anti-rigging accessory and ACCUWIK were weighed, weighed and the .Lol weight recorded. The anti-rotation accessory was then fitted with a cell separation module and the threaded positioning part was screwed into the cell separation module at the bottom. The numbers stamped both on the cell separation modules and on the positioning handles were removed. used to establish the position with a compile loop corresponding to 1 mm of vertical movement. The anti-rotation handle was first placed just below the VIVID platform on the cell separation module to allow the 18 mm VIVID cut to be adhered, The 18 mm fpi adhesive ring is used to attach the 18 mm VIVID to the cell separation module using the margins as a guide. The threaded positioning device was then used to position the ACCUWIK at distances of 0.5 mm, 0 mm, 0.25 mm and 0.5 mm from the bottom surface of the VIVID membrane, The distance, d, is defined as the distance between the lower surface of VIVID and the upper surface of the handle until irritation (lower surface of ACCUWIK) (SEE FIG. 10). The ascending direction is negative. 125 pL of whole blood (50% hematocrit) was added to VIVID. 50% hematocrit was achieved by rotating the whole blood samples in a centrifuge (4,000X) for 10 minutes, transferring all the plasma to a new tube and adding the same volume of cellular components to the plasma sample. Blood with a sufficient 50% hematocrit was prepared accordingly. when the same blood count was used for all tests. The sample was left in filtration for 10 minutes. The VIVID membranes are then removed and appropriately stripped in a re® Lp already from the biological riseu. The anti-rotation piece was also removed and weighed with a scale. Opeso was reg is trade. The ACCUWIK membrane was then discarded in a biologically reciprocal pie rite. The volume of plasma collected was calculated by selecting the difference between pre- and post-weight weights and according to the density of the plasma. The density of pJasma 'was estimated at 1025 kg Ar. The plasma recovery was then calculated using the volume of plasma collected by the total amount of plasma in the original sample, which in this case was 57.5 pL, and multiplied by 100.
    The distances of 0 mm and 0.25 mm produced the most altar recovery of 52 pL and 53 pt. However, at a distance of 0.25 the variability (standard deviation) was 5.9111 compared to 0.9411 at 0 mm. At a distance of 0.5 mm, the ACCUWIK membrane (thickness - 0.38 - 0.53 mm) may no longer be in contact with the VIVID membrane and therefore cannot complete another Plasma Lanto as a result of the dim i nu i g.l & na capillary forces. At a distance of 0.5 mm [indicative that ACCUWIK is completely compressed against VIVID}, Col observed high plasma recovery; however, plasma recovery was not as high as in the other two distances. This is most likely due to the large drop in the emptied volume caused by the compression of the material's fibers, although the present invention is not limited to any specific action required and a census of the mechanism of action is not necessary. paraprati cur the presence presence. Based on exposing the elements conducted during the development of the present invention, a 0 mm scale was used in Ex-emp 7 os 13 ^ 13. Example 7 Effects of Sample Variables
    Experiments were conducted during the. development of modalities of the present invention to determine the effect of blood volume on plasma collection with methods, systems and devices of the present invention. The volume of blood obtained from a specific puncture device depends on the main factors: physical / mechanical (eg, the puncture device itself), bioldgenic (eg, the thickness of the skin, the individual's hematocriuo, the individual's weight, the amount and potency of the clotting factors in the individual's blood, etc.) and the process (eg, the puncture position, the ability to maintain adequate contact between the puncture and the skin during the process punishment, etc.). Although the volume of blood collected is variable, in some modalities, methods, systems and devices must collect plasma volume constants to quantitatively measure plasma constituents, such as the actual HIV-1 plasma levels.
    Usually in practice, a plasma aliquot is collected for testing after being separated from the blood by a central leak. However, this complies with a procedure; it usually requires the use of two devices: 1) a centrifuge to separate blood and plasma and 2) equipment: accurate to measure plasma; and l.has a suitable procedure for use in the field. Therefore, in some embodiments, the present invention provides a simple, low-cost device for removing a fixed amount of plasma, regardless of the volume of the original sample (e.g., the amount of blood obtained from a digital needle).
    In some embodiments, the present invention provides an absorbent membrane (e.g., ACCUWIK) that is sized to collect a fixed amount of plasma, leaving the plasma residue on the VIVID GR membrane. In some embodiments, the present invention provides a low-cost device that allows direct blood collection on a filter paper, and not on a Liar cap tube.
    Experiments were conducted during the development of modalities of the present invention to demonstrate that a device of the presence invention would collect a fixed amount of plasma regardless of the amount of blood added, Fresh blood from a human individual was used for the study. A known volume of blood with a 45% hematocrit level was added to the device. The device was left to stand for 1U minutes, and the volume of plasma collected was measured. 100, 125, 150 and 175 p-L of blood were added to the devices, and the plasma volume was. measured (See FIG. Hl). Addition. less than H 25 pL resulted in the collection of 3 9.6 gL of plasma, with an efficiency of 88.1%. Therefore, a minimum volume of 125. pL is necessary to collect 50 µg of plasma. The gradual addition of blood volumes up to 175 ph. did not result in an increase in the volume of plasma collected. The maximum eticienuia of plasma collection is 94.6% and obi Ida with the addition of 12.5 pL of plasma, which is run with a progressive mother in the volume of blood. Figure 11 also shows the variation of the variability of plasma plasma levels; o Cyclic Threshold for 600 sets of IltV-1 / mL of blood. This was estimated by the use of the HIV-1 standard curve (y -3.33x i 3.1.5) obtained by purifying different concentrations of virus in the plasma and amplifying it. In some embodiments of the present invention, 600 copies / ml is the lower limit of detection, therefore, the effect of changing the volume on the CT would be the highest with that number. The CT changes from 25.49 to 25.56 when the blood added to the blood-plasma device changes from 125 pL to 175 pL. Is.Lo is satisfactorily within the accepted range of +0.5 specified by our test device. Experiments were conducted during the development of modalities of the present invention in which the VIVID Plasma Separation GR membrane was dimensioned according to the manufacturer's protocols, and sizes different from the PALL VIVID membrane were tested, while maintaining the optimized contact pressure between the VIVID membrane and the ACCUWIK and the size of the ACCUWIK membrane (See FIG. 12). Increasing the size of the membrane reduces the efficiency of purification. In all test groups, the plasma collection module was separated from the separate 10 minute module after adding blood to the blood plasma unit.
    In some modalities, experiments were conducted to determine the effect of hemat.6cr.Lto on plasma collection. Hematberite is the proportion, by volume, of blood that consists of blood cells. A normal level of haematperiLia would be significant according to gender, age, sailing status etc. Once a fixed volume of blood has been determined, thereafter at the hematocrit levels, the amount of plasma available for purification is increased. They also affect aviscosi, blood quality, therefore interfering in the flow rate through a device. The PALL VIVID Plasma Separation GR membrane is a highly asymmetric membrane that separates hemaclas from plasma by filtration by size exclusion. Therefore, high levels of hematocrit can clog the membrane resulting in inadequate plasma separation. Experiments were carried out to determine the effect of such changes in the hematocrit on the volume of plasma collection from a fixed volume of whole blood. A device of the present invention was prepared as described previously, and blood from an individual tinnitus was centrifuged at 3,500 rpm for 20 minutes to sepate the cells. The plasma levels were adjusted to create blood samples with 5 different hematocrit levels between 15-60%. 125 IJL of the blood sample was then added to the blood-plasma module and the volume of plasma collected in the plasma module was measured. The volume changed from 53.2 pL to 47.6 pL at the five different hematocrit levels, corresponding to an estimated change in CT of 0.16 to a concentration of 600 copies of HIV-1 / mL, estimated using the standard HIV-1 curve ( y = - 3.33x + 31.5) obtained by purifying different virus concentrations in the plaque or its amplification. The results show how much the blood-plasma module can successfully succumb to different levels of hematocrit without any obstruction of the VIVID Plasma Separation GR membrane. The volume of collated plasma declines according to the increase in. hematocrit dec.orr.onte of reduced amount of plasma in the sample- In a urn. hematocrit level of 60%, 125 pL of blood contains only 50 μl of plasma. Therefore, the plasma collected is also low. At a 15% hematocrit level, available plasma is 106.25 pL. However, only 51.3 µL are used in the plasma module once. that the PALL ACCUWIK membrane is saturated and does not allow more plasma flow to the module. Example 8 Fresh Whole Blood Separable
    Experiments were conducted during the development of modalities of the present invention to demonstrate the effectiveness of the plasma separation and collection module to efficiently separate whole blood plasma obtained from different human patients. Like blood and an extremely complex matrix, its variability. Age between people can affect their rheological and flow properties 15 aLraves of the membrane, which in turn would affect the purification efficiency of the sample. Fresh blood was obtained from Northwestern Memorial Hospital for this study. The blood sample was homogenized by placing the tube in a rotary mixer 20 for 2 minutes. The hematocrit of blood samples was measured by rotating 30 µl of the blood in Light Cycler PCR Tubes (F. Hoffmann-La Roche Ltd., Basel, Switzerland) at 3,000 rpm. for 10 minutes. Hema 1.6c r i L © was measured. 125 pl. of blood were added and plasma toil collected. The plasma separation and collection module has successfully plotted the different blood samples (SEE FIG. 14). The average volume of plasma collected is 125 pl. of plasma was ■ 19.5 pL, 0 standard deviation of the volume of plasma collected and 3.3 b pl.i. It was observed how the separation separation was 30 s significantly better when fresh blood was collected and decreased as the blood sample aged. All tests were carried out with blood samples collected by venous pung in a vacuum tube containing anticoagulants.
    Experiments were conducted during the development of present invention modalities to test the efficiency of separation and collection of blood plasma with high leukocyte cell count. 53 pL of plasma were successfully collected from 125 pL of whole blood, suggesting that a high cell count of leukocytes does not obstruct the membrane. The test was performed using the protocol © described above. Example 9 Determination of Viral Load of Whole Blood
    Experiments were conducted during the development of modalities for the presence of the invention, in which a determined viral load was used to demonstrate the feasibility of the plasma separation system for all-purpose applications. This Objective was to separate blood cells from plasma, concentrate the plasma on a smaller ACCUWIK disk and then use it to extract viral RNA from I1IV-1, for purification and for PCL based lossless detection. siqnif 1 caL i.va in the RNA in the modules. of separation or collection. 1 * 1 asthma was separated from 1 whole blood using: “- if a provision of the prescribed invention: or standard laboratory contrifugation,
    Samples for processing technique were prepared by separating blood cells and blood plasma by means of leakage at 3,500 rpm for 10 minutes. The proportion of plasma ® cells to be mixed to reconstitute a blood sample with 45% hematocrit percentage® was calculated. Plasma was inoculated with HFV-1 virus obtained from Northwestern Memorial Hospital (1.50'0 eopias / pL plasma supply concentration) 5 to produce a raped concentration of 300, 60 and 12 Oopia.s / pL Respectively, the blood was reconstituted using that plasma and cells by mixing them in a proportion of 45% cells and 55% plasma.
    For samples procured by a device of the present invention, 125 µl of the above blood was added to the VIVID membrane of the plasma collection and separation module and left to stand for 10 minutes to allow for plasma separation. VIVID has been severely removed. The ACCUWIK was inserted into a centrifugal primer tube (Tube 1) 15 containing 400 µL of Ambion lysis buffer that includes Ambion Blnding / Lysis Concentrate, isoprop alcohol) T, ico and Ambion Carrier RNA.
    For samples- processed by cent. In the standard laboratory leak, blood was leaked at 3,000 rpm for 10 minutes to separate the plasma from the cells. 50 pl. of plasma were then added to ACCUWIK of the same size as used for the plasma separation and collection module. The ACCUWIK is inserted in a tuft of r.:crc;:er L r 1 leak containing Use Ambion cap which includes Ambion 25 B i nding / Lysis COT iGKnt rate, ail cool isoprop) 1 i Co and Ambion Ca r r i and r RNA.
    All samples are then mixed by vo-rLex for 2 minutes. 10 pl. Spherical mixtures containing Ambion PMRs and LnLonslf1cator from the Ambion linkage were added and not vortexed for 4 minutes. 200 pL of the solution was pipetted and transferred to another carefully removed Pickpen-1 Magnetic (SunrDiego, CA) tube (Tube by means of Science: from one product also placed in another tube. Remaining in the first tube was eliminated. APMPs. sample in Tube 2 was then processed using the PF method (VERFIG.large of the Tubocarrier sample and mixed into 1.1). 5 0 pL2 was added during 4 minutes of capping the chamber, using the same method as the chamber: smaller than the I PFos cartridge Aqueous fluids were covered as shown in Figure 15. The automated system added the 'PIMP's; for 2 minutes the cover using the external magnet and the new aggregate deodorizer to contain the elution cap removed from the elution cover. Quantification of HIV-1 cargaviral was performed using bHIV-1 Amplification Kit in volumes of the albumin reaction (T 9531; Fisherseries bovinaSigma) and 0.2% Scientific) ampli f Leagao were (Sunnyvale, CA) .quadrup'la.suggesting that Abbott Rea (Time (Abbott Molecular25 pL with addition) Since (B8667, Sigma), 159 mM of Tween 20 (28320; pL each load (Planes, IL) 0.2 mg / mL of trehaloseThermocommodus Mode I O'-in Cepheidviral foieonduz Age II in quadruple.
    Efficiency of BCR dplasplasma was observed, productive semt. ransportchemacias queini.well PCR.The results obtained from blood centrifugation and separation using the plasma separation and collection modules were identical, demonstrating that there was no significant loss in viral particles in the filling membrane, Example 10 Storage of PMP in the Membrane Matrix
    Experiments were conducted during the development of modalities of the present invention to demonstrate the feasibility of storing PMPs in the ACCUWIK membrane. In some embodiments, the plasma containing the HIV-1 virus is separated towards the ACCUWIK membrane through the VIVID membrane. This membrane is subsequently inserted into the tube containing the lysis plug, whereby the virus undergoes lysis and the viral RNA binds to PMPs. The viral RNA diffuses out of the paper towards the lysis plug containing the PMPs. To facilitate: the process, the solution containing the paper is stirred. A portion of the RNA is lost in the paper as a result of absorption by the membrane fibers. In some embodiments, to minimize the loss of RNA in the filter membranes and eliminate the need for agitation, PMPs are pre-distributed at ACCUWIK. When a sample containing viral RNA is added to the paper, the virus and the PMPs are both located in the matrix. porous of the ACCUWIK membrane. With the addition of lysis reagents, the viral RNA can immediately isolate the PMPs without diffusing out of the paper. The membrane is therefore used as a storage matrix. of rcagenie and subosoquenl and RNA capLura. For further processing of the sample, the magnetic particles are extracted from the paper by means of a low cost magnet or permanent electromagnet. This process eliminates the need for shaking, heating and centrifuging associated with RNA extraction from the filter. In addition, it provides a convenient location for reagent storage in service areas near the top.
    In experiments conducted during the development of modalities of the present invention, BMPs from the Arab i Ph Magmax kit were added as alquota in a micro.centrif.uga tube and placed on a magnetic © support to collect the PMPs. The liquid was removed and the PMPs. were suspended again in Ambion Binding Enhancer. For each test sample, 10 in. of PMPs and 10 ylj of Ambion Binding Enhanuet were used. A 5 µl solution containing 4% BSA and 0.4% Triton-X is added. The solution was added to a 12 mm disc of ACCUWIK Ultra with an area sufficient to hold 50 µl of sample and allowed to air dry for 3. days at room temperature. Plasma was inoculated with HIV-1 virus obtained from Northwestern Memorial. Hospital. (Concentration of the supply of 1,500 qdpi / pl. Of plasma) to produce a concentration of 300, 60 and 1'2 copies / pL, respectively. The air-dried ACCUWIK Ultra was inserted into a microcentrituga tube (Tube 1) © 50 pL of the sample. of plasma were added to the paper. 4 μl of Ambion lysis buffer was added to the solution, which was left to stand for 6 minutes. The particular magnets were removed from Tube 1 using one: Pi.ckpeh-1 Magnetic tool (Sunrise Sejonoe Products Inc., San Diego, CA) and I. Transferred to 400 pL of Ambion Wash Buffer-1 in the Tube 2. Sample in Tube 2 was then processed using the TPF method (SEE FIG. 15.). The sample from Tube 2 was added to the large chamber of the cartridge and mixed for 4 minutes by means of an automated system. 50 µl of elution buffer was added ■ as a whole to the smaller chamber of the IPF cartridge and the liquid aqued dots were covered with CHILLOUT wax liquids (Biorad laboratories; SEE FIG. 15). The automated system aggregated the PMPs for 2 minutes by means of an external magnet and moved the aggregate from the lysis plug to the elution plug. The volume of dluigS containing the PMPs was heated to 55 ° C for 10 minutes to elute the. RNA. PMPs were added and removed from the cap. elution. Quantification of HIV-1 viral load was performed using the Abbott RealTime HIV-1 Amplification Reagent Kit [19] (Abbott Molecular, Des Plaines, IL) in 25 pt solution volumes with the addition of 0.2 mg / ml of serum albumin. bovine (138661; Sigma), 150 mM Lrealose (T9531; Sigma) and 0.2% Tween 20 (28320; Pierce Thermo Fisher Scientific) and 5 pL as a model. Amplification reactions were performed on Cepheid SmartCycler II (Sunnyvale, CA).
    For flask samples, plasma was inoculated with HIV-1 virus obtained from Northwestern Memorial Hospital (concentration of the supply of 1,500 copies / piL of plasma) to produce a race concentration of 300, 60 and 12 copies / kti respective i va to. 50 'pL of the plasma sample containing different concentrations of the HIV-1 virus was added to a microcentrifuge tube (Tube J). 400 uL. of Amb i on lysis buffer were added to Tube 1 and mixed by vertex for 30 seconds®. 20. gL of mixture of spheres containing 10 µL of PMP.s and 10 µL of binding device were added and mixed by mixing for 4 minutes. The magnetic particles were removed from Tube 1 using a Pickpen-1 Magnetic tool (Sunrise Science Products Inc., San Diego CA) and transferred to 400 pL of Ambion Wash Buffer-1 in Tube 2. The sample in Tube 2 was then processed using the IFF method (SEE FIG. 15). The sample from Tube 2 was added to the larger chamber of the cartridge and mixed for 4 minutes by an automated system. 50 µl of elution buffer was added as an aliquot to the smaller chamber of the IFF cartridge and the two aqueous liquids were covered with CHILLOUT wax liquids (Laboratories Biorad; SEE FIG. 15):. The automated system aggregated the PMPs for 2 minutes using an external magnet and moved the aggregate from the lysis plug to the elution plug. The elution buffer containing the PMPs was heated to 55 ° C for 10 minutes to elute the RNA. PMPs were aggregated and removed from the elimination buffer.
    Quantification of HIV-1 viral load was performed using Abbott Real! Ime HIV-1 Arnpiif: ica L..1 on Reagent Kit [IB] (Abbott Molecular, Des Plaines, IL) in reaction volumes of 25 pL with the addition of 0.2 mg / ml of bovine serum albumin (138667, Sigma;), 150 mM trehalose (T9531; Sigma) and 0.2% Tween 20 (28320; Pierce Thermo Fisher Scientific) and 5 pl, as a model. Amplification Reactions Cepheid SmartCycler IL (Sunnyvale, CA). fri i conducted in doubl. IgaLas. As HIV-1 negative Go.ntrdle, other samples were used.
    The viral RNA was purified from 50 µl of plasma inoculated with HJV-1 virus. The purified RNA was amplified using the Abbott R.ealTlme HIV-1 Amplification Kit. PCR efficiency of 107.7% was observed (SEE FIG. 17), indicating that PMPs can be stored in the membrane with no efficiency in the cap. of RNA and can be readily extracted by downstream processing. The minimum amount of viral RNA on the filter membrane is 6 (See FIG. 18). Example 11 Plasma Separation
    The lateral flow system has been used in the detection and diagnosis of HIV. Current laboratory technologies are based on the combined and simultaneous detection of HIV core proteins (p24) and anti-HIV and spedic antibodies directed against HIV trans-human proteins. Antibodies against such proteins appear consistently during seroconversion of HIV-infected individuals and remain throughout the course of infection. In the first 2 months after birth, HIV-positive infants have a progressive viral load, but are seropositive as a result of the inheritance of maternal anti-HIV antibodies, which Lorna has ineffective existence tests for. In addition, HIV-negative infants may be seropositive due to the same inheritance of anti-HIV antibodies in Lemos. Detection of HIV in infants requires targeting the p24 protein from the HIV nucleus as the marker principle for datccgSo with the aim of verifying the true status of infection regardless of maternal health inheritance.
    Experiments were carried out during the development of the present invention modalities for realizing the usefulness of separating blood by means of a membrane for the purpose of adjusting the plasma collection on a collection pad. Plasma separation devices were assembled using an 18 mm VIVID GT 'membrane and an 8 mm Ahlstrom 142 fiberglass base collection pad or 6 mm Pall A / D. The collection pad was compressed 0.5 mm into the VIVID GF membrane. The VIVID membrane was ionized to accommodate a fixed volume of blood, up to .125 pL, and the Ahlstrom / Pa .11 membranes were sized to accommodate a fixed volume of 50 U.L. The VIVID membrane was pretreated with a solution containing 1% BSA protein, 0.5% sucrose and 0.1% Tween 20. 100 µl of the pretreatment solution was added to each VIVID membrane and allowed to air dry for 24 hours at room temperature before use. Samples of fresh human blood of 100 pl or 125 pF, by volume were added to the devices and the plasma collection volume was calculated based on the mass differences of the collection membrane before and blood separation depots.
    For 1.25 p.L of whole blood, 48 ± 2 pl. of plasma were collected with an average collection time of 8 ± 2 minutes by both Ahlstrom and Pall membranes. For 100 pL of whole blood, 37 ± 2 pL of plasma were collected with an average collection time of 5 ± 2 minutes, also across both Ahlstrom and Pall membranes. Such data indicate that this membrane-based system can be used offetively to syntonize and adjust the collection of plasma from a sample. of whole blood.
    Experiments were conducted during the development of modalities of the present invention to determine the extent of analyte losses not specified for the collection pad of the. VIVID and Ahlstrom membranes, where the protein amphigene p24 of. HIV was diluted in human plasma and passed through separation devices. Separation devices were assembled as described above. For a separation device, the VIVID membrane is not pre-read. rated with BSA / sucrose / 'Tween. In a second, the solution of 0.1% BSA, 0.5% sucrose and 0.1% Tween 20 was used as a pre-treatment as previously described. A third sample was prepared with 50 pL of plasma containing 1 ng / mL of analyte p24 added directly to the Ahlstrom collection pad. Finally, 50 μl of plasma containing analyte p24 was added directly to a tube: containing an analysis buffer, and this reference reaction had the composition and the labeled. Each reaction tube was pulse vortexed to thoroughly mix the reaction. Each tube was then heated to 95 ° C for 4 minutes. Each tube was then allowed to cool to room temperature (2.3 ° C) for 4 minutes. Biotinylated morioc 'on.al antibody against a p24 epitope was added to each reaction vessel. After 1 minute, 13.5 pL of carbon conjugate coated with a monoclonal antibody against a second p24 epitope was added to the reaction. After brief mixing, a test strip containing a neutral grav i d.f.na test line to an anti-mouse control line was added to the reaction. The test tapes were qualified using a camera after being dried by subtracting the Sinai background piano from the test line. Only marginal recovery losses (eg, non-specific connection) of p24 were observed in the separation device fitted with an untreated VIVID membrane (VDK EIG. 19). For- all other samples. An equivalent result such as that of the reference mix and the conduction reactions was obtained, indicating no significant loss of the analyte signal in relation to the VIVID membrane: as for the collection pad. Ahlstrom. Example 12 Plasma Separation and Detection of p24
    Experiments were. performed during the development of modalities of the present invention to evaluate the recovery of analyte p.24 inoculated in whole blood and recovered from plasma after separation. Blood samples were reconstituted from a fresh blood sample with two hematocrit values (Ht) (50% and 35% Ht) that reflect the clinical range found in the humafia species. Volumes of each sample were used (125 µl, 50% Ht and 100 µl 35% Ht) so that the total volume of plasma available for recovery was the same (60. pL, of which a maximum of 50: pL can be absorbed by the collection membrane). Separation devices were assembled and whole blood re-constituted (125 pL of 50% Ht and 100 pL of 35% Ht) was applied to the separation devices, and plasma collected from the pad of each was subjected to endorsement ; action for reading the signal., an additional Teferencla reaction was promoted by means of. Wed! 50 pl. of plasma containing 1 ng / ml of the p21 analyte were added directly to the anaerobic buffer with the rotated composition and determination. The amount of analyte recovered from the application of samples of tannin blood with Ht of 35% as well as 50%: it is comparable The composition and the determination of the reference reaction (SEE ELG. 20), Indicating that this device Plasma separation can be used to successfully recover protein analytes from whole blood samples without any detectable analyte losses that occur through the separation of the Gelular sariguine phase.
    The same experiment was carried out by assembling separation devices fitted with the Pall A / I membrane in place of Ahlstrom 142 and reconstitution of whole blood with 35% Ht inoculated with 1 ng / ml of analyte p24. Beginning earlier, an additional referral reaction was performed whereby 50 µl of plasma containing 1 ng / ml of p24 analyte was added directly to the Use aria buffer with the labeled composition and measurement. The amount of analyte recovered from the application of blood samples with 35% Ht containing p24 is comparable to the composition and determination of the reference reaction (SEE. FIG. 21). This result confirms that the ability to collect and elute p24 analyte in a membrane is applicable to different compositions of the collection membrane. Example 13 Storage of Analysis Reagents in the Plasma Separation Module
    Experiments were carried out during the development of modalities of the present invention to demonstrate the feasibility of storing analysis reagents in the plasma collection membrane. In some modalities, Lateral flow analysis uses a specific concentration of detergents, namely, SOS or NP-40, to allow viral breakdown (HIV) and analyte gap of the Cp24 analyte) of samples infected with IILV . Thus, for a plasma sample of 50 µL, 100 µL of PH.S buffer containing 0.2% SDS and 0.67% NP-40 are preferentially necessary to break the virion and recover p24. Experiments were carried out in which 10 UL of a ten times concentrated detergent solution sypcimerite (2.0% SDS and 6.7% NP-40) were applied to a collection pad and allowed to dry for 24 hours at room temperature. environment. They were then mounted in a separator and plasma collected from a 100 µl whole blood sample, inoculated with 1 ng / mL of the p24 analyte. The result was measured against a reaction of composition reference and determination of the solution-phase type with an equivalent volume (50 pL) of plasma. The result of the analysis of the desiccated detergent on the pad was comparable to the composition reference solution and has a solution-phase type action, indicating that the drying of the analysis reagents is feasible and compatible in the collection pad membrane (SEE FIG. 22) ■ Example 14 Detergent and Analyte Desiccation on Cushion
    Eeitas aval iagoos through which 10 pt of ten times concentrated detergent solution supply (2.0% SDS and 6.7% NP-40) were applied to a collection pad and allowed to dry for 24 hours at room temperature. environment. Then, 50 µl of plasma with or without 1 ng / ml of the p24 analyte was added to the pad or allowed to dry for 24 hours at room temperature. The samples were then replenished with 125 µl of PBS buffer, mixed by shaking the tube containing the cushion and plug and analyzed. The result of the detective analysis / p24 desiccated no. cushion toi comparable to the reaction of composition reference and determination of the solution-phase type, indicating the desiccation of the analysis agents as well as the protein. p24 used as power! reference standard (positive and negative controls) for an analysis that is feasible and compatible in the collection pad membrane (SEE FTG. 23). Example 15 Device with Shell-type Filter Module
    In some embodiments, the present invention provides a filter module with a shell-like pattern (SEE FIG. 24) which is closed by folding and locking the upper half by locking tabs (SEE FIG. 23). In some embodiments, a shell-type filter module comprises a slot for a collection tube designed to accommodate the width of the collection test strip. In some embodiments, the slot entrance has both the chan.Trad.as edges and a filled bottom edge to help guide a collection module into the shell without interference. In some embodiments, the depth of the gap is projected in such a way that there is an overlap between the diaphragm. collection and separation filter (SEE FIG. 30). In some embodiments, a shell-type filter module comprises locking algae (eg left and right locking algae) which. permilom lock the top half closed (see FIG. 25). In some embodiments, the shell is anchored by an operator by pressing on the lower half of the locking algae to rotate and release the upper half of the shell. In some modalities, a hinge is integrated into the shell design. In some embodiments, a plastic shell component is made of polypropylene. In some modalities, the hinge provides the ability to open and close the shell thousands of times, although the real process requires only flex.So for a short time. In some embodiments, a shell-type filter module comprises one or more bags (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 ..., 20). In some embodiments, the bags serve to maintain uniform thickness of the molded part, which prevents warping and depression marks, such as those caused by uneven freezing. In some embodiments, the wall thickness for the bags is uniform. In some embodiments, a shell-type filter module comprises a flap. In some modalities, - when the upper half of the shell is locked using the locking algae, the flap fits with a fold in the collection modifies. (SEE FIG- 31). Once fitted and locked in place, the test-collection tape cannot leave the shell by itself unless the operator releases the top half of the shell. In. in some embodiments, the shell-type filter module comprises lock protections (eg 4 lock protectors). In some embodiments, lock guards prevent the top half of the shell from sliding back and forth once it has been locked in place by the locking algae. In some embodiments, the lock guards. they fit with the edges of the locking algae (SEE FID. 25). In. In a modal modalities, the shell type fl! trb module comprises an energy manager. Km some modalities ,. an energy manager facilitates the connection of the separate filter with the shell by means of an uassasonic welding medium (SEE LUGS. 26-27). In some imadul i Jades, the triangular energy manager is designed so that its reading is approximately the same as the thickness of the separation line. In some embodiments, the last single-sided weld creates a pseudo-ring barrier as the plastic of the energy manager deepens and flows into the separation filter. The pseudo-ring type barrier facilitates greater separation performance. In some modalities, the membrane is connected using thermal sealing, laser welding, adhesives, etc. In some embodiments, a shell-type filter module comprises a rear wall. In some embodiments, the back wall of the slot serves as a breakpoint for positioning the collection modules. When the collection module is positioned to make contact with the rear wall, the fold automatically aligns with the shell .ab (see FIG. 31}.
    In some embodiments, the shell filter module is configured to connect to, function with and / or integrate with a separation filter. In some embodiments, a shell-type filter module is integrated with a separation filter (eg, Pall Vivid) (SEE FIG. 26). In some embodiments, the soup filter is any suitable filter, as described here and in other locations. In some embodiments, the separation filter is connected to the upper half of the shell by means of wol'd-agcm ulbrassdnica using energy qerenciadd.r. In other embodiments, the separation filter is switched on using thermal sealing, laser welding, adhesives, etc. In some embodiments, when the separation filter is connected to the upper half of the shell and closed, the separation filter is essentially flush with the lower half of the shell (SEE FTC. 27, the right) - In some modalities, ultrasonic welding .sonica provides the ability of the molded plastic of the energy manager to merge, flows and re-joins the separation filter, creating a pseudo-ring clay that further facilitates the separation by .1. i rn: i Excessive volume of flow inside the separation filter that reduces the amount of plasma captured by the collection membrane.
    In some embodiments, the shell filter module is configured to connect to, operate with and / or integrate with a collection module (eg plasma collection module, plasma collection module, etc. .). In some embodiments, a collection module comprises a collection membrane (eg, Pall A / D). In some modalities, the collection membrane and the component that Collects it absorbs the specimen of interest, J's to 6, plasma. In some embodiments, a collection module comprises three main components - substrate test tape, adhesive and collection membrane. In some embodiments, the substrate test tape is a thin plastic sheet that is flexible enough to be bent and punctured, but also sufficiently inflexible to hold the adhesive and the collection membrane. In some modalities, the collection module is factory.Leads using Leenoloqia standard current used in tape.s for testing and similar lateral flow. In some embodiments, the test line fold serves to align the flap with the shell (See FIG. 31). In some embodiments, proper fold orientation ensures that the flap fits exactly with the right angle to prevent the string from coming out of the shell. In some embodiments, the tape 6 is also perforated to allow the operator to easily pull it over the collection module and to cut it into two separate pieces, if desired for the analysis procedure. In some embodiments, a double lace adhesive (3M, St Paul, MN) compatible with diagnosis and used to join the collection membrane to the substrate test tape. In some embodiments, the adhesive is compatible with diagnosis (that is, it does not contain or release any of the biochemical inhibitors that would interfere with biochemical analysis) and is double-sided to safely bond to both types of substrate (substrate test tape) and collection membrane). In some embodiments, the adhesive resists rigorous exposure to high temperatures (<100 ° C) or liquids (plasma, saline solutions, buffered with 150 mM concentrations and detergents, such as SDS and NP-40, with conceriLragbes S 0.15% and 0.5%, respectively i vameril.e). In some embodiments, the collection membrane (eg Pall A / D) is sized to be the same width as the collection tape and matches the geometry of the adhesive on the other axis.
    In some embodiments, to separate the blood sample, the plasma collection module (PCM) is inserted into the filter module (SEE FIG. 29). With the filter module in the open position, the PCM is slid independently by the slit and aligned with the rear wall, ensuring that the collection membrane is electronically separated with the separation Tilt to the flap, with the fold, from the line to LesLe of the PCM substrate. In some ways, the height of the slit in the filter module is projected so that there is a replacement between the separation filter and the collection membrane. In some embodiments, the overlap facilitates compression between the separation filter and the collection membrane. In some embodiments, cotpressure facilitates blood separation (SEE FIG. 30).
    In some embodiments, when the PCM is inserted into the filter module and the shell is closed, the shell tab (SEE FIG. 24) fits with the fold in the substrate testing tape (SEE FIG. 28). In some embodiments, the flap compresses smoothly into the PCM, ensuring that it is held firmly in the PCM. In some embodiments, the rear edge of the flap fits with the edge of the fold (SEE FIG. 31), which prevents the PCM from sliding out of the shell.
    This embodiment of the present invention is used to perform separation of a blood sample (SEE FIG, 32). The operator distributes the volume of blood prescribed in the separation filter. After waiting a few minutes, the separation filter allows the passage of blood plasma, but not other blood components, to the collection membrane in the collection module. Con La to Intimo between the separating filter and the collecting membrane allows passage of blood plasma and facilitates the separation of blood (See FIG. 30). Once the separation is complete, the operator presses the bottom half of the left locking algae and pulls the tabs outward to remove the top half of the filter module. At this stage, the fold of the. collection is no longer attached to the tab, and the collection module can be removed by the user. The collection and retrieval module of the I itro module. Either or both: ih6dul.os can be used for subsequent LesLc> processes. In some embodiments, the patient's CD4 count can be monitored using the filter module that contains the cellular components of the blood. In some cases, the collection module, which contains the blood plasma component, can be used to determine viral load using PCR or related biochemical reaction. For example, PMRs separate from or housed in the PCM can be used for extraction of viral RNA from the blood plasma and eluting directly into the cap for PGR related processing. In some circumstances, PCM can be used in a side flow system for the detection of anti-HIV antibodies, p24 protein etc. In some circumstances, the filter module and / or the PCM may be transported to a secondary institution for any of the biochemical processes mentioned above. REFERENCES
    All publications and patents mentioned in the preceding DESCRIPTION and / or listed below are hereby incorporated by reference. Various modifications and variations of the described compositions and methods of the invention will be evident to those with practical experience without deviating from the purpose and principle of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific specimens. In fact, several modifications of the models described to conduct the Invention that are obvious to those with experience in the relevant fields aim to be within the scope of the present invention. The following references are incorporated by reference in their entirety:
    Kelso D, Sur K, Parpia Z: Barriers for facilitating biological reactions. U.S. Pat. App. No. 20030246782, October 1, 2009.
    Brenner T, Haeberle S, Zengerle R, Ducree J: Continuous centrifugal separation of whole blood on a disk. SPECIAL PUBLICATION-ROYAL SOCIETY OF CHEMISTRY 2004, 296: 566-568.
    Haeberle S, Brenner T, Zengerle R, Ducree J: Centrifugal extraction of plasma from whole blood on a rotating disk. Lab on a Chip 2006, 6: 776-781.
    Kang J, Cho H, Kwak S, Yoon D, Kim T: Novel particle separation using spiral channel and centrifugal force for plasma preparation from whole blood. SPECIAL PUBLICATIONROYAL SOCIETY OF CHEMISTRY 2004, 296: 614-616.
    Madou M, Lee L, Dauner! S, Lai S, Shih C: Design and fabrication of CD-like microfluidic platforms for diagnostics: microfluidic functions. Biomedical Microdevices 2001, 3: 245-254.
    Toner M, Irimia D: Blood-on-a-chip. Annual Review of Biomedical Engineering 2005, 7: 77-103.
    Luo W, Yang H, Rathbun K, Pau Cop, Ou CoY: Detection of Human Immunodeficiency Virus Type 1 DNA in Dried Blood Spots by a Duplex Real-Time PCR Assay. J Clin Microbio / 20C5, 43: 1851-1857.
    Blattert. C, Jurischka R, Tabhan I, SchoLh A, Kerth P, Menz W: Separation of blood in microchannel bends. In .: 2004.
    Kondo A, K.il.ajima M: Multilayered integral chemical analysis element tor the blood. Fuji Photo Film Co., Ltd., 1981.
    Vogel P, Braun H, Berger D, Werner W: Process and compositionLion for separating plasma or serum from whole blood. (USPTO ed.: Boehringer Mannheim GmbH; 1984.)
    Vogel P, Braun H, Berger D, Werner W: Device for separating plasma or serum from whole blood and analyzing the same. Boehringer Mannheim GmbH; 1989.
    Baumgardner J, Loewen M: Blood separation media and method for separating plasma from whole blood. Ahlstrom Filtration, Inc .; 1993.
    Loewen MD, Baumgardner JS: Development of a nonwoven filler for seperation of serum from whole blood for medical diagnostic applications. Tappi Journal 1996, 79: 183-184
    Simon LD: Blood seperation medium usefil for seperating erythrocytes from whole blood comprises non-woven web of fibrillated and non-fibrildated syntheticstaple fibers. Pp. 6: SIMON L D (SIMO-Individual) AHLSTROM MT HOLLY SPRINGS LLC (AHLS Non-standard): 6.
    Hillman R, Gibbons I; Blood separation device comprising a filter and a capillary flow pathway exiting the filter. Biotrack, Inc .; 1988.
    Suzuki H, Ho CM: A magnetic force driven chaotic micro-mixer. Pp. 40-43; 2002: 40-43.
    Chien 5>: Red cell deformability and its relevance to blood flow. Annual Review of Physiology 1987, 49: 177-192.
    BaskurL OK, Mei sei man ILJ: Blood rheology and hemodynamics. Seminars in Thrombosis and Hemostasis 2003, 29: 435-450.
    Benson K: MCAT review. Emory University 1999: 141-199.
    Huang S, Sall bore J, Tang N, Luk KC, Hackett Jr, J, Swanson P, Cloherty G, Mak WB, Robinson J, Abravaya K: I'hermodynamiGally modulated partially double-stranded linear D.NA probe design for homogeneous real -time PCR. Nuc Acids Res. 2007, 35 :: el01.
    Mylonakis E, Paliou M, Rich J: Plasma viral load 5 testing in the management of HIV infection. Am Fam Physician 2001, 63: 483-490.
    Dineva MA, Mahilum-Tapay L, Lee H: Sample preparation: -a challenge in the development of point-of-care nucleic acid-based assays for reset roe limited settings. Analyst10 2007, 132: 1193-1199.
    权利要求:
    Claims (16)
    [0001]
    1. Blood plasma filtration method characterized by the fact that it comprises: providing a device that comprises a collection membrane configured to allow the passage of blood plasma more quickly than the cellular components of blood and a plasma collection module configured to extract blood plasma into it by means of capillarity, in which the plasma collection module accommodates a fixed volume of plasma, with the plasma collection module in direct contact with the collection membrane, applying a blood sample to the collection membrane, in which blood plasma is collected in the plasma collection module, where the fixed volume of plasma in the plasma collection module is greater than the volume of plasma collected in the plasma collection module; and separate the plasma collection module from the collection membrane.
    [0002]
    2. Method, according to claim 1, characterized by the fact that said extraction of blood plasma through the collection and passive membrane.
    [0003]
    3. Method, according to claim 2, characterized by the fact that said extraction does not require electrophoresis, centrifugation or more than atmospheric pressure.
    [0004]
    4. Method, according to claim 1, characterized by the fact that said collection membrane accommodates a fixed volume of said blood sample.
    [0005]
    5. Method, according to claim 1, characterized by the fact that said fixed volume of said plasma collection module is less than said fixed volume of said collection membrane.
    [0006]
    6. Method, according to claim 1, characterized by the fact that said fixed volume of said plasma collection module is independent of the volume of said blood sample.
    [0007]
    7. Method, according to claim 1, characterized by the fact that said fixed volume of said plasma collection module is independent of the hematocrit of said blood sample.
    [0008]
    8. Method, according to claim 1, characterized by the fact that it also includes storage of said plasma in said plasma collection module.
    [0009]
    9. Method, according to claim 1, characterized by the fact that it still comprises the insertion of the plasma collection module into a module for analysis.
    [0010]
    10. Device for separating plasma from whole blood characterized by the fact that it comprises: a filter module comprising (i) a collection membrane configured to allow only the passage of blood plasma, and (ii) a collection module comprising a plasma collection module, in which the filter module and the collection module are connected so that the collecting membrane and the plasma collecting module are in close contact to extract the blood plasma from the collecting membrane by means of capillarity; and where the plasma collection module is separable from the collection membrane, where the plasma collection module accommodates a fixed volume of plasma that is greater than the volume of plasma extracted into the plasma collection module
    [0011]
    11. Device, according to claim 10, characterized by the fact that the said collection membrane accommodates a fixed volume of whole blood.
    [0012]
    12. Device, according to claim 10, characterized by the fact that said fixed volume of said plasma collection module is less than said fixed volume of said collection membrane.
    [0013]
    13. Device, according to claim 10, characterized by the fact that the said collection membrane comprises a plasma separation membrane based on asymmetric polysulfone.
    [0014]
    14. Device, according to claim 10, characterized by the fact that said collection membrane comprises collection pad based on fiberglass.
    [0015]
    15. Device, according to claim 10, characterized by the fact that it comprises paramagnetic particles in the referred plasma collection module.
    [0016]
    16. Device, according to claim 10, characterized by the fact that it comprises analysis reagents in the referred plasma collection module.
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    同族专利:
    公开号 | 公开日
    US20120024788A1|2012-02-02|
    WO2012015926A3|2012-05-03|
    CN103229053A|2013-07-31|
    WO2012015926A2|2012-02-02|
    CN103229053B|2015-06-24|
    BR112013002064A2|2016-05-24|
    ZA201301455B|2014-04-30|
    US10386356B2|2019-08-20|
    US9816979B2|2017-11-14|
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    JPS587332Y2|1978-06-06|1983-02-08|
    DE3029579C2|1980-08-05|1985-12-12|Boehringer Mannheim Gmbh, 6800 Mannheim|Method and means for separating plasma or serum from whole blood|
    US4816224B1|1980-08-05|1992-03-10|Boehringer Mannheim Gmbh|
    US4753776A|1986-10-29|1988-06-28|Biotrack, Inc.|Blood separation device comprising a filter and a capillary flow pathway exiting the filter|
    US5423989A|1988-05-19|1995-06-13|Chemtrack, Inc.|Plasma forming device|
    NL8800796A|1988-03-29|1989-10-16|X Flow Bv|METHOD FOR THE CHEMICAL ANALYSIS OF BODY FLUID COMPONENTS, AND A TESTING DEVICE AND TEST PACKAGE FOR SUCH ANALYSIS.|
    US5186843A|1991-07-22|1993-02-16|Ahlstrom Filtration, Inc.|Blood separation media and method for separating plasma from whole blood|
    US5589399A|1994-10-21|1996-12-31|First Medical, Inc.|System and method for plasma separation and measurement|
    US5569399A|1995-01-20|1996-10-29|General Electric Company|Lasing medium surface modification|
    WO1996035952A1|1995-05-09|1996-11-14|Smithkline Diagnostics, Inc.|Devices and methods for separating cellular components of blood from liquid portion of blood|
    US6036659A|1998-10-09|2000-03-14|Flexsite Diagnostics, Inc.|Collection device for biological samples and methods of use|
    DE60043049D1|1999-12-28|2009-11-12|Arkray Inc|Blood test device|
    JP3808393B2|2002-03-28|2006-08-09|富士写真フイルム株式会社|Blood test unit and blood test apparatus|
    US6755802B2|2002-05-06|2004-06-29|Beckman Coulter, Inc.|Whole blood sampling device|
    EP1580551B1|2002-11-19|2013-02-20|Sekisui Medical Co., Ltd.|Plasma or serum separation membrane and filter apparatus including the plasma or serum separation membrane|
    US7228973B2|2003-05-23|2007-06-12|Ahlstrom Mt. Holly Springs, Llc|Nonwoven fibrous media especially useful for the separation of blood constituents|
    DE102004027422A1|2004-06-04|2005-12-29|Boehringer Ingelheim Microparts Gmbh|Device for receiving blood and separating blood components|
    CN101568636A|2006-11-02|2009-10-28|威腾技术公司|Cartridge for conducting diagnostic assays|
    CA2716950C|2008-02-29|2017-07-04|Northwestern University|Barriers for facilitating biological reactions|
    US8148071B2|2008-09-30|2012-04-03|Northwestern University|Methods and compositions for isolating nucleic acid|
    WO2010129302A1|2009-04-28|2010-11-11|Innovative Laboratory Technologies, Inc.|Lateral-flow immuno-chromatographic assay devices|
    BR112013002064B1|2010-07-27|2020-11-24|Northwestern University|DEVICE AND METHODS FOR BLOOD PLASMA FILTERING|BR112013002064B1|2010-07-27|2020-11-24|Northwestern University|DEVICE AND METHODS FOR BLOOD PLASMA FILTERING|
    EP2845001B1|2012-05-03|2016-12-14|Qualigen, Inc.|Whole blood analytic device and method therefor|
    US9938568B2|2013-07-26|2018-04-10|General Electric Company|Ligase-assisted nucleic acid circularization and amplification|
    US9644232B2|2013-07-26|2017-05-09|General Electric Company|Method and device for collection and amplification of circulating nucleic acids|
    JP6138260B2|2013-09-09|2017-05-31|株式会社日立製作所|Cell separation / recovery membrane, culture sheet and culture apparatus using the same, and cell separation / recovery method|
    JP6566319B2|2013-12-03|2019-08-28|国立大学法人 東京大学|Separation unit, separation method, fluid device, composite fluid device and kit|
    WO2015095491A1|2013-12-19|2015-06-25|The Trustees Of The University Of Pennsylvania|Plasma separator apparatus and associated methods|
    EP3083056A4|2013-12-20|2017-08-02|Spot on Sciences, Inc.|Whole blood separation sampling apparatus|
    US20170226467A1|2014-07-31|2017-08-10|Govind Rao|Microscale bioprocessing system and method for protein manufacturing from human blood|
    JP6715236B2|2014-08-01|2020-07-01|シーメンス・ヘルスケア・ダイアグノスティックス・インコーポレーテッドＳｉｅｍｅｎｓ Ｈｅａｌｔｈｃａｒｅ Ｄｉａｇｎｏｓｔｉｃｓ Ｉｎｃ．|Pressure assisted plasma separation|
    US9950321B2|2014-08-04|2018-04-24|General Electric Company|Device for separation and collection of plasma|
    EP3294133B1|2015-05-14|2019-09-04|General Electric Company|Method and kit for separation and collection of plasma|
    US10335078B2|2014-08-04|2019-07-02|General Electric Company|Device for separation and collection of plasma|
    US10111610B2|2014-11-04|2018-10-30|Wainamics, Inc.|Microscale plasma separator|
    CN104437128A|2014-11-10|2015-03-25|苏州蔻美新材料有限公司|Nanometer magnetic hemodialysis membrane and preparation method thereof|
    WO2016093999A1|2014-12-12|2016-06-16|The Trustees Of The University Of Pennsylvania|Fluid separator for point of care molecular diagnostics|
    WO2016158139A1|2015-03-31|2016-10-06|ソニー株式会社|Electrical characteristic measurement device, electrical characteristic measurement method, blood condition analysis system, and electrical characteristic measurement program for computerizing said method|
    US10906042B2|2015-07-21|2021-02-02|Gattaco Inc.|Automatic plasma separation and metering|
    CN108473932A|2015-09-09|2018-08-31|集联健康有限公司|For sample collection, stabilisation and the system of preservation, method and apparatus|
    KR20180080241A|2015-11-02|2018-07-11|바이오파이어 다이어그노스틱스, 엘엘씨|Sample preparation for difficult sample types|
    US20170212112A1|2016-01-25|2017-07-27|General Electric Company|Integration of sample separation with rapid diagnostic tests for improved sensitivity|
    US10983119B2|2016-01-25|2021-04-20|General Electric Company|Device for rapid diagnostic tests to detect antigens with improved sensitivity|
    CA3015627A1|2016-02-24|2017-08-31|Partnership For Clean Competition|Multilayer device for separating blood components and uses thereof|
    AU2017268787A1|2016-05-25|2018-12-06|Monolythix, Inc.|Processing blood samples to detect target Nucleic Acids|
    CN106053193A|2016-06-02|2016-10-26|北京师范大学|Method and device for collecting and storing biological components in urine by filter|
    CN208808491U|2017-01-10|2019-05-03|集联健康有限公司|A kind of device for from subject's collection fluid sample|
    WO2018175169A1|2017-03-20|2018-09-27|Wainamics, Inc.|Small volume self-metered blood separation device|
    CN108732001B|2017-04-24|2021-03-05|光宝电子有限公司|Plasma and blood cell separation device and separation method thereof|
    TWI644691B|2017-04-24|2018-12-21|天亮醫療器材股份有限公司|Blood plasma and blood cell separation device and separating method thereof|
    EP3619532A1|2017-05-02|2020-03-11|Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V.|Plasma/serum separator device and methods using the same|
    WO2019016080A1|2017-07-17|2019-01-24|Boehringer Ingelheim Vetmedica Gmbh|Modified filter membrane and the use thereof|
    JP2021527212A|2018-06-12|2021-10-11|ナウダイアグノスティック、インコーポレイテッド|Devices and methods for collecting plasma|
    WO2020068910A1|2018-09-25|2020-04-02|The Trustees Of Columbia University In The City Of New York|Blood sample processing and nucleic acid amplification systems, devices, and methods|
    WO2020251849A1|2019-06-12|2020-12-17|Siemens Healthcare Diagnostics Inc.|Plasma separation and sample metering device and kits and methods of use related thereto|
    CN110339876B|2019-06-27|2021-04-30|昆明理工大学|Tumor cell screening microfluidic device and method based on droplet deposition|
    WO2021026261A1|2019-08-05|2021-02-11|Auer Precision Company, Inc.|Microfluidic passive plasma separation device and method|
    WO2021076846A1|2019-10-18|2021-04-22|Tasso, Inc.|Plasma separation devices and related methods|
    法律状态:
    2018-04-10| B06F| Objections, documents and/or translations needed after an examination request according art. 34 industrial property law|
    2019-07-16| B06T| Formal requirements before examination|
    2020-08-11| B09A| Decision: intention to grant|
    2020-11-24| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 27/07/2011, OBSERVADAS AS CONDICOES LEGAIS. |
    优先权:
    申请号 | 申请日 | 专利标题
    US36815610P| true| 2010-07-27|2010-07-27|
    US61/368,156|2010-07-27|
    PCT/US2011/045541|WO2012015926A2|2010-07-27|2011-07-27|Devices and methods for filtering blood plasma|
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