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WO1997032213A1 - Trousse d'analyse du sang - Google Patents

Trousse d'analyse du sang Download PDF

Info

Publication number
WO1997032213A1
WO1997032213A1 PCT/US1997/002889 US9702889W WO9732213A1 WO 1997032213 A1 WO1997032213 A1 WO 1997032213A1 US 9702889 W US9702889 W US 9702889W WO 9732213 A1 WO9732213 A1 WO 9732213A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
membrane
blood
limited portion
absorbent pad
Prior art date
Application number
PCT/US1997/002889
Other languages
English (en)
Inventor
Howard Greenspan
John Colanduoni
Original Assignee
Majesco Biologicals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Majesco Biologicals, Inc. filed Critical Majesco Biologicals, Inc.
Priority to AU21912/97A priority Critical patent/AU2191297A/en
Publication of WO1997032213A1 publication Critical patent/WO1997032213A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing

Definitions

  • This invention relates generally to appara t us for testing blood including tests for blood type or viral infections, such as HIV. More specifically, this invention relates to the use of membranes in blood testing apparatuses.
  • Blood testing is normally performed in a laboratory setting requiring several steps for each type of test to be performed. While the present invention is directed toward two basic type of blood tests, blood type testing and testing for viral infections, other applications will be apparent to those skilled in the art.
  • Blood typing is performed by detecting the type of antigen that is contained within a person's blood. Specifically, a person with A type blood has the A antigen in their blood cells; people with B type blood have the B antigen in their blood cells; people with AB type blood have both the A antigen and the B antigen in their blood cells; and people with 0 type blood have neither the A nor the B antigen.
  • Another process utilizes size exclusion chromatography.
  • a mixture of blood and antibody is placed in a test tube on top of a layer of gel.
  • the tube is then placed in a centrifuge. If the sample contains red cells with the specific antigen on their surface, the cells and antibody will clump together and will not be able to be centrifuged through the gel.
  • the antigen or reactant could be disposed on an underside or an opposing side of a membrane. Blood then could be forced to proceed through the membrane before reacting with the antigen or other reactant. The remaining components of the blood could be thereafter absorbed or washed away leaving the blood cell-reactant aggregate disposed against the underside of the membrane. Because membranes are clear, the result could be an easy-to-read indicator such as a line or a dot. Further, separate antigens or reactants could be placed at discrete locations on the membrane to provide multiple indication in one test kit.
  • both A type and B type as well as RH positive antibodies could be placed on discrete areas of the membrane so that one test apparatus could provide positive indications for A, B, AB, O, RH positive and RH negative blood.
  • Such a system would not require the use of a centrifuge or excessive handling by a technician.
  • the invention is an improved apparatus for testing blood, such as testing blood type and testing for viral infections.
  • One apparatus made in accordance with the present invention includes a backing sheet for accommodating a membrane on one surface section of the backing sheet and an absorbent pad on a second surface section of the backing sheet.
  • the membrane and absorbent pad are in a juxtaposed relationship with one another.
  • antibodies corresponding to A, B and RH positive blood cells are arranged in discrete areas of the membrane.
  • One preferred method is to place a horizontal "stripe" of antibody corresponding to each blood type in a parallel relationship.
  • the backing sheet, absorbent pad and membrane are then cut into strips.
  • the purpose of the absorbent pad is to absorb liquid as it flows through the membrane. When the blood cells react with an antibody or other reagent, the aggregation is too large to pass through the membrane and a red stripe appears where the antibody or reagent was placed on the membrane.
  • a second and related embodiment features a membrane mounted onto the top of an absorbent pad.
  • the absorbent pad includes a hole or aperture disposed in the middle of the pad.
  • the membrane/ pad combination are then accommodated in a housing which includes an opening or access hole disposed above the portion of the membrane covering the hole or opening in the absorbent pad.
  • the appropriate antibodies or reagents are previously placed in discrete areas of the membrane portion which is disposed over the hole in the pad. Blood is then applied directly to the top of the membrane and subsequently washed away with a buffer solution. Blood cells which react with the antibodies or reagents form an aggregation or bright red area after the remaining components of the blood are washed away with the buffer solution.
  • a membrane having approximately a 20 micron pore size is preferable for use with both blood testing apparatuses described above.
  • Red blood cells are approximately seven to eight microns in diameter. Thus, unreacted red blood cells will pass through the 20 micron membrane. However, once the red blood cells react with an antibody or other suitable reagent to produce an aggregate, the aggregate cannot pass through the 20 micron pore size. As the remaining components of the blood are washed away w ch a buffer solution, the aggregate combination is left behind :o provide a bright red indicator on the under surface of the membrane. Because the membranes are transparent, a clear visual indication is provided.
  • a membrane with a smaller pore size, such as 3-6 microns is provided so that the blood and reagents will bind to the surface of the membrane.
  • Still another object of the present invention is to provide an improved apparatus for testing viral infections in human blood. Another object of the present invention is to provide an improved method for testing blood.
  • Figure 1 is a schematic diagram illustrating a positive reaction between an antibody and a red blood cell with the specific antigen and a negative reaction between an antibody and a red cell without the specific antigen;
  • Figure 2 is an illustration of the prior art method of using size exclusion chromatography in testing blood
  • Figure 3 is a top plan view of a plastic backing sheet used in one blood testing apparatus made in accordance with the present invention
  • Figure 4 is a top plan view of the plastic backing sheet shown in Figure 3 with a membrane applied to a lower surface section thereof,-
  • Figure 5 is an illustration of the backing sheet and membrane shown in Figure 4 further with an absorbent pad attached to the backing sheet in a juxtaposed relationship with respect to the membrane;
  • Figure 6 is an illustration of the embodiment shown in Figure 5, cut into strips and illustrating the reactions for A- , A+, B- , B+, AB-, AB+, O- and 0+ blood types;
  • Figure 7 is a side view illustrating the attachment of a membrane to an absorbent pad for a second embodiment made in accordance with the present invention.
  • Figure 8 is a top plan view of the absorbent pad first shown in Figure 7;
  • Figure 9 is a side sectional view illustrating the membrane and absorbent pad shown in Figure 7 contained within a housing.
  • Figure 10 is an illustration of eight blood testing apparatus shown in Figure 9 indicating A- , A+, B-, B+ AB-, AB+, O- and 0+ blood types.
  • FIG. 1 an antibody 11, a red blood cell 12 and a red blood cell 13 are shown at the left.
  • the red blood cell 12 includes an antigen which will react positively with the antibody 11 which will result in an aggregation, shown generally at 14.
  • the aggregation 14 is a positive reaction or a positive indication that the red blood cell 12 is of the blood type specific to the antibody 11.
  • the antibody 11 will not react with the red blood cell 13 because the red blood cell 13 lacks the specific antigen.
  • the combination of the antibody 11 and the red blood cell 13 is a negative reaction or lack of clumping, indicated generally at 15.
  • the aggregation or agglutination shown at 14 is useful in size exclusion chromatography as shown m Figure 2.
  • a size exclusion chromatography method is illustrated in Figure 2.
  • a test tube 16 is filled with a gel 17.
  • a mix type of red blood cells and a specific antibody are placed on top of the gel 17 at the area indicated generally at 18.
  • the test tube 16 is then placed in a centrifuge (not shown) . If no reaction takes place between the antibody and the red blood cells, the red blood cells shown at 20 will centrifuge downward toward the lower end of the test tube below the gel, indicated generally at 19.
  • the test tube shown m the middle of Figure 2 indicates a negative reaction, because the red blood cells 20 were able to pass through the gel 17 during the centrifuge process.
  • red blood cells shown in the test tube 16 at the far right of Figure 2 reacted with the antibody and formed an aggregation 21 which was not able to pass through the gel 17 during the centrifuge process.
  • the red blood cells in combination with the antibodies are shown at 21, disposed on top of the gel 17.
  • the aggregation was unable to pass through the gel and therefore the area disposed at the bottom 19 of the test tube 16 remains clear.
  • the size exclusion chromatography process shown m Figure 2 is inconvenient for several reasons. Specifically, only one antibody or reagent may be used at a time. Thus, several tests are required to determine blood type which requires the use of several test tubes. Further, a centrifuge is required.
  • Figure 3 is an illustration of a backing sheet 22 which, as shown in Figure 4, accommodates a membrane 23.
  • the dotted lines 24, 25 which pass through the membrane 23 are intended to indicate the placement of antibodies on either the top side or underside of the membrane 23.
  • Figure 5 illustrates the placement of an absorbent pad 26 on top of the backing sheet 22 and m a juxtaposed relationship with respect to the membrane 23. As fluid passes through the membrane 23, it is absorbed into the pad 26.
  • the apparatus 27 shown in Figure 5 can then be cut into strips 28a tnrough 28h as shown in Figure 6.
  • the membrane 23 has been coated with three specific antibodies -- the specific antibody for A type blood, the specific antibody for B type blood and the specific antibody for RH+ blood. As illustrated to the left of the strip
  • the A type, B type and RH+ antibodies are coated on the membrane in spaced apart parallel stripes.
  • the use of the three different antibodies enables the test strips 28 to indicate eight different results.
  • the test strip 28a gives a positive indication for A- blood.
  • the test strip 28b is a positive indication for A+ blood.
  • the test strip 28c gives a positive indication for B- blood; the test strip 28d gives a positive indication for B+ blood.
  • the test strips 28e and 28f gives positive results for AB- and AB+ blood respectively.
  • test strips 28g, 28h give positive results for 0- and
  • test strips 28a through 28h as shown m Figure 6 can be used as follows: the blood samples are collected in a standard method with any type of anticoagulant to stop clotting. Approximately five microliters of blood is then added to a 12mm by 75mm test tube along with approximately 95 microliters of buffer containing 1% BSA, 50mM sodium phosphate ph 7.4, with 0.1 percent sodium azide . The strip 28 is then added to the tube with the membrane 23 on the bottom and the solution is allowed to migrate up the strip 28 to the absorbent pad 26. The results can then be observed within one to four minutes.
  • a second embodiment of the present invention is illustrated in Figure 7. Specifically, a membrane 30 is placed on top of an absorbent pad 31 as shown in Figure 7. As shown in Figures 7 and 8, the pad 31 includes a hole or aperture 32 over which the membrane 30 is placed. As shown in Figure 9, the membrane 30 and pad 31 are contained within an outer housing that includes a top 33 and bottom 34. Use of the apparatus 35 is further illustrated m Figure 10.
  • Each membrane 30 includes three different antibodies, placed m discrete and separate positions, as illustrated by the kit 35f which indicates AB+ blood.
  • the kit 35a indicates a positive reading for A- blood; the kit 35b indicates a positive reading for A+ blood.
  • the kits 35c, 35d indicate positive readings for B- and B+ blood respectively.
  • kits 35e, 35f indicate positive readings for AB- and AB+ blood respectively.
  • the kits 35g, 35h provide positive readings for 0- and 0+ blood respectively.
  • the kit 35 is used as follows. Fifty microliters of blood is placed on the top of the membrane 30. The blood is then allowed to sit for sixty seconds. The membrane is then washed with lmL of the above-described buffer solution to remove the unreacted blood cells. The results as shown m Figure 10 are then interpreted. A red dot on the left hand side denotes A type antigen. A red dot on the right hand side denotes B type antigen. A red dot m the upper center portion of the membrane 30 indicates RH+ .
  • the preferred membrane is comprised of mixed esters of cellulose.
  • the preferred pore size is approximately 20 microns. However, the pore size may vary.
  • One preferred memDrane is sold under the HIFLOW-SXTM which is sold in strips that are twelve inches wide, having a thickness of 115-155 microns.
  • the top 33 and bottom 34 may be manufactured from plastic, such as PVC.
  • the backing sheet 22 may also be manufactured from plastic. Instead of utilizing a 20 micron membrane, a membrane having a smaller pore size, such as three microns may be utilized. If a 3 micron membrane is utilized, the antibodies should be applied to the top or front surface of the membrane.
  • a 20 micron membrane may be utilized alone, without being fabricated into a strip form, such as the strips 28 as shown m Figure 6 or the kits 35 as shown in Figure 10.
  • ten microliters of blood may oe mixed with twenty microliters of antibody solution with fifty microliters of buffer. This mixture may then be pipetted directly onto the membrane. A bright red spot will appear in positive samples as the agglutinated antibody red cell complex will not be able to pass through the twenty micron pore membrane. In a negative sample, the red cells pass through the membrane and no clumping is observed.
  • membrane m accordance with the present invention will be useful for other types of testing, other than blood-type testing. Specifically, the invention and methods of using the invention will be useful for the detection of viral infections, such as HIV. Further, the present invention may be utilized to detect any type of a blood condition that may be otherwise detected by agglutination or aggregation.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention porte sur une trousse améliorée d'analyse du sang et son procédé d'utilisation. Ladite trousse peut être fournie sous plusieurs formes dont une forme en bande dans laquelle une membrane (30) et un coussinet absorbant (31) sont juxtaposés sur une couche substrat. Une partie de l'un des côtés de la membrane (30) est traitée par des anticorps réagissant avec les antigènes des groupes sanguins A, B et RH+. Dans une autre variante, la membrane est placée sur un coussinet absorbant (31) à l'intérieur d'un boîtier. Un trou ou une ouverture (32) est pratiqué dans le coussinet (31) sous une partie de la membrane (30). Des parties distinctes de la membrane sont traitées par des anticorps spécifiques des antigènes des groupes sanguins A, B et RH+. Si les antigènes sont disposés du côté inférieur de la membrane, on utilise une membrane présentant des pores d'environ 20 microns; si les antigènes sont disposés au dessus de la membrane ou sur sa face supérieure, on utilise une membrane présentant des pores plus petits, de 3 à 8 microns.
PCT/US1997/002889 1996-03-01 1997-02-27 Trousse d'analyse du sang WO1997032213A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU21912/97A AU2191297A (en) 1996-03-01 1997-02-27 Blood testing kit

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US60970596A 1996-03-01 1996-03-01
US08/609,705 1996-03-01

Publications (1)

Publication Number Publication Date
WO1997032213A1 true WO1997032213A1 (fr) 1997-09-04

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Application Number Title Priority Date Filing Date
PCT/US1997/002889 WO1997032213A1 (fr) 1996-03-01 1997-02-27 Trousse d'analyse du sang

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AU (1) AU2191297A (fr)
WO (1) WO1997032213A1 (fr)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000020862A1 (fr) * 1998-10-02 2000-04-13 Abp Diagnostics Ltd. Procede et appareil destines a la detection in vitro de substances multiples
WO2005003787A1 (fr) * 2003-07-08 2005-01-13 Inverness Medical Switzerland Gmbh Dispositif et procede de detection d'agglutination de particules
WO2005005991A1 (fr) * 2003-07-09 2005-01-20 Medion Diagnostics Gmbh Dispositif et procede de determination simultanee d'antigenes de groupes sanguins
WO2007029250A1 (fr) * 2005-09-06 2007-03-15 Inverness Medical Switzerland Gmbh Procede et appareil de configuration d'un substrat buvard
WO2006051548A3 (fr) * 2004-11-15 2007-05-24 Inverness Medical Switzerland Procede, systeme et dispositif de typage sanguin
FR2931244A1 (fr) * 2008-05-13 2009-11-20 Biosynex Sarl Dispositif de detection d'au moins un element contenu dans une solution sanguine
EP1644735B1 (fr) * 2003-07-09 2011-09-21 Medion Diagnostics AG Dispositif pour tests sur membrane a ecoulement lateral (lateral flow)
CN102288746A (zh) * 2011-07-12 2011-12-21 英科新创(厦门)科技有限公司 一种血液筛查联合检测卡
WO2012159275A1 (fr) * 2011-05-26 2012-11-29 Siemens Aktiengesellschaft Système de détermination de groupe sanguin
CN103502819A (zh) * 2011-04-21 2014-01-08 西门子公司 血型测定装置以及检测血型的方法
WO2021175239A1 (fr) * 2020-03-04 2021-09-10 重庆大学 Carte de test de système à multiples groupes sanguins automatique et procédé de test

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4379847A (en) * 1979-09-19 1983-04-12 American Hospital Supply Corporation Suspending medium for immunologic reactions
US4459361A (en) * 1982-06-04 1984-07-10 Angenics, Inc. Ligand assay with one or two particulate reagents and filter
US4552839A (en) * 1983-08-01 1985-11-12 Syntex (U.S.A.) Inc. Determination of analytes in particle-containing medium
WO1988008534A1 (fr) * 1987-04-27 1988-11-03 Unilever Plc Immuno-analyses et dispositifs pour les realiser
US4851210A (en) * 1986-05-22 1989-07-25 Genelabs Incorporated Blood typing device
US4943522A (en) * 1987-06-01 1990-07-24 Quidel Lateral flow, non-bibulous membrane assay protocols
US5213963A (en) * 1988-10-12 1993-05-25 Biotest Aktiengesellschaft Procedure for finding and identifying red cell antibodies by means of the solid phase method

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4379847A (en) * 1979-09-19 1983-04-12 American Hospital Supply Corporation Suspending medium for immunologic reactions
US4459361A (en) * 1982-06-04 1984-07-10 Angenics, Inc. Ligand assay with one or two particulate reagents and filter
US4552839A (en) * 1983-08-01 1985-11-12 Syntex (U.S.A.) Inc. Determination of analytes in particle-containing medium
US4851210A (en) * 1986-05-22 1989-07-25 Genelabs Incorporated Blood typing device
WO1988008534A1 (fr) * 1987-04-27 1988-11-03 Unilever Plc Immuno-analyses et dispositifs pour les realiser
US4943522A (en) * 1987-06-01 1990-07-24 Quidel Lateral flow, non-bibulous membrane assay protocols
US5213963A (en) * 1988-10-12 1993-05-25 Biotest Aktiengesellschaft Procedure for finding and identifying red cell antibodies by means of the solid phase method

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000020862A1 (fr) * 1998-10-02 2000-04-13 Abp Diagnostics Ltd. Procede et appareil destines a la detection in vitro de substances multiples
WO2005003787A1 (fr) * 2003-07-08 2005-01-13 Inverness Medical Switzerland Gmbh Dispositif et procede de detection d'agglutination de particules
US8053226B2 (en) 2003-07-09 2011-11-08 Medion Diagnostics Ag Device and method for simultaneously identifying blood group antigens
WO2005005991A1 (fr) * 2003-07-09 2005-01-20 Medion Diagnostics Gmbh Dispositif et procede de determination simultanee d'antigenes de groupes sanguins
KR101214620B1 (ko) 2003-07-09 2012-12-21 메디온 디아그노스틱스 아게 혈액형 항원을 동시에 결정하는 장치 및 방법
EP1644735B1 (fr) * 2003-07-09 2011-09-21 Medion Diagnostics AG Dispositif pour tests sur membrane a ecoulement lateral (lateral flow)
WO2006051548A3 (fr) * 2004-11-15 2007-05-24 Inverness Medical Switzerland Procede, systeme et dispositif de typage sanguin
WO2007029250A1 (fr) * 2005-09-06 2007-03-15 Inverness Medical Switzerland Gmbh Procede et appareil de configuration d'un substrat buvard
FR2931244A1 (fr) * 2008-05-13 2009-11-20 Biosynex Sarl Dispositif de detection d'au moins un element contenu dans une solution sanguine
CN103502819A (zh) * 2011-04-21 2014-01-08 西门子公司 血型测定装置以及检测血型的方法
WO2012159275A1 (fr) * 2011-05-26 2012-11-29 Siemens Aktiengesellschaft Système de détermination de groupe sanguin
CN103765221A (zh) * 2011-05-26 2014-04-30 西门子公司 血型测定系统
CN102288746A (zh) * 2011-07-12 2011-12-21 英科新创(厦门)科技有限公司 一种血液筛查联合检测卡
WO2021175239A1 (fr) * 2020-03-04 2021-09-10 重庆大学 Carte de test de système à multiples groupes sanguins automatique et procédé de test

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