WO1989007763A1 - Procede servant a detecter la presence d'anticorps dans le sang et bande de membrane et batterie de membranes utilisees lors de la realisation dudit procede - Google Patents

Procede servant a detecter la presence d'anticorps dans le sang et bande de membrane et batterie de membranes utilisees lors de la realisation dudit procede Download PDF

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Publication number
WO1989007763A1
WO1989007763A1 PCT/DK1989/000025 DK8900025W WO8907763A1 WO 1989007763 A1 WO1989007763 A1 WO 1989007763A1 DK 8900025 W DK8900025 W DK 8900025W WO 8907763 A1 WO8907763 A1 WO 8907763A1
Authority
WO
WIPO (PCT)
Prior art keywords
membrane
strip
antigenes
antibodies
coated
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/DK1989/000025
Other languages
English (en)
Inventor
Cosma Francis De Souza
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ANCOS AS
Original Assignee
ANCOS AS
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 ANCOS AS filed Critical ANCOS AS
Priority to BR898907241A priority Critical patent/BR8907241A/pt
Priority to FI903964A priority patent/FI903964A0/fi
Publication of WO1989007763A1 publication Critical patent/WO1989007763A1/fr
Priority to NO90903535A priority patent/NO903535L/no
Priority to DK190390A priority patent/DK190390A/da
Anticipated expiration legal-status Critical
Priority to BG092796A priority patent/BG92796A/bg
Ceased legal-status Critical Current

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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
    • G01N33/53—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
    • 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
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G01N33/548—Carbohydrates, e.g. dextran

Definitions

  • the present invention relates to a method for detecting the presence of antibodies in blood by immunoblotting- technique (Western blot) using a membrane coated with screened antigenes corresponding to the antibodies to be detected.
  • the invention further relates to a membrane strip and a membrane battery for use when carrying out the method.
  • Antigenes can e.g. be separated and detected by gel electrophoresis, e.g. SDS.
  • gel electrophoresis e.g. SDS
  • SDS gel electrophoresis
  • the advantages of the so-called blotting technique include i.a. antigene concentration on the surface of a membrane in immobilised state, so that the individual separated antigenes are easily accessible for further identification.
  • An analysis method based on immunoblotting-technique can be used for characterization of both antibodies and antigenes.
  • antibodies which can be detected in this way include all kinds of retrovirus, such as HIV-I, HIV-II, HTLV-I and HTLV-II.
  • retrovirus such as HIV-I, HIV-II, HTLV-I and HTLV-II.
  • Other examples are malaria, schistosomasis, filariasis, African trypanosomiasis and laishmaniasis.
  • allergy producing substances can be detected.
  • the normal immunoblotting-technique employs blood samples which are pretreated for isolating serum.
  • the isolated serum is contacted with a membrane in the form of a strip, width typically 4-5 mm, and length 120 mm, and which is coated with screened antigenes corres- ponding to the antibodies to be detected. After suitable incubation time the membrane is removed and tested for positive reaction. Testing may be by any known method/ such as colouring.
  • the present invention is a further development and simplification of the known immunoblotting-technique, and involves significantly shortened analysis time. Furthermore, the analysis is made on whole blood in stead of serum, e.g. a blood sample which has been transferred onto a carrier and has coagulated.
  • the invention relates to a method of the kind stated in the introductory part of claim 1, being characterized in that a carrier with the blood sample which has coagulated after application onto the carrier, is introduced into a container together with the membrane in the form of a strip coated with the screened antigenes and an elution and incubation liquid, and that the membrane after a preset incubation time is removed and tested in a way known per se.
  • the present invention it is possible to carry out elution and incubation in one single step, which entails a significant simplification and shortening of the analysis time. Contributing to the obtained shortening of the analysis time is also the fact that with the method according to the invention it is possible to use a membrane in the form of a strip with extremely small dimensions. Preferred dimensions of the membrane strip used are: width 1.1-1.8 mm and length 10-100 mm. This considerably reduces the price of the process, and also the time necessary for elution and transfer of anti ⁇ bodies onto the membrane is shortened considerably, e.g. from 3-4 hrs. to 1-1 1.5 hrs.
  • the preferred membrane strip for use when carrying out the method according to the invention is a polyvinyl- idendifluoride foil, cut into strips of the stated dimensions, i.e. preferably a width of 0.1-1.8 mm and a length of 10-100 mm, e.g. 70-75 mm.
  • Examples of other suited membrane materials are nitrocellulosis or poly- amide.
  • Each strip can be marked with an identification code, such as a sticker with a journal number or a bar code directly on the film strip.
  • test For carrying out the test use can be made of an arbitrary container capable of containing the membrane and the carrier with the blood sample as well as elution and incubation liquid. If a large number of tests are to be carried out simultaneously it is expedient to use a block with a number of cells into which a corresponding number of membranes and carriers with blood sample can be introduced.
  • FIG. 1 is a perspective view of a block according to the invention
  • Fig. 2 is a cross-section of a part of the same block
  • Fig. 3 is a perspective view of one of the two lids of the block.
  • Fig. 4 is a cross-section similar to that of Fig. 2, in which the membrane, carrier with blood sample and elution and incubation liquid have been introduced.
  • Fig. 5 is a membrane battery with 15 membrane strips, coated with similar or different antigenes,
  • Fig. 6 is a membrane battery of the same type as shown in Fig. 5, each membrane strip, however, being divided into 3 sections.
  • Fig. 7 is a side-view of the membrane battery shown in Fig. 6,
  • fig. 8 is a side and partly sectional view of a test block with a number of chambers suited for introduction of the membrane battery shown in Fig. 7,
  • Fig. 9 is a top view of the test block shown in Fig. 8,
  • Fig. 10 is a section through a test block chamber along the line a-a in Fig. 8, and
  • Fig. 11 is a carrier strip for a number of pieces of filter paper with blood samples, suited for introduction into the chambers of the test block shown in Figs . 8-10.
  • the test block consists of a plate 1 of an autoclavable material, such as POM.
  • a number of parallel through- going holes 2 has been bored in the plate, into which holes through-going tubes 3 projecting in excess of both sides of the plate 1, have been inserted.
  • the thickness of the plate 1 is 30 mm.
  • the diameter of the holes 2 is 10 mm- 2 - * . 80 mm long tubes, projecting 25 mm in excess of both sides of the plate 1, have been introduced into the holes, so that the openings 6 of the tubes are free of the surface of the plate, and each tube end is provided with an annular siliconelastomer seal 9.
  • the openings 6 can be closed with two lids 4 ad 5 provided with recesses 7 corresponding to the tube openings.
  • Fig. 4 shows a crosse-section of a tube into which a piece of filter paper with blood sample 8 and a membrane strip 10 have been introduced.
  • the two tube openings 6a, 6b are closed with lids 4, 5.
  • each hole 2 of a block as shown in the drawing is introduced a strip of a polyvinylidendifluoride film provided with a reinforcement rib, which film in a manner known per se has been coated with screened HIV antigenes.
  • the strips have a length of 70 mm and a width of 1.6 mm.
  • the strips used are each labelled with stickers provided with printed on journal number. Then pieces of filter paper with applied blood samples from the persons to be tested are cut to dimensions of 5 x 5 mm.
  • a membrane battery as shown in Fig. 5 comprising a number of membrane strips 3 being attached at a binding site 4 to flaps 2 on a carrier strip 1 of a water-repelling plastic material.
  • the membrane strips 3 are coated with similar or different antigenes corresponding to the antibodies to be detected.
  • the membrane battery is introduced in a block with the same number of holes or through-going borings and at the same interspaced distance as the membrane strips, corresponding to a row in the block shown in Figs. 1-4. Elution and incubation liquid and pieces of filter paper with dried up blood samples from the persons to be tested are introduced simultaneously into the said holes or through-going borings.
  • Figs. 6 and 7 show a membrane battery consisting of a holder or rib 11 with a.number of flaps 12 provided with a corresponding number of membrane strips 13 of a plastic material, such as polyvinylindendiflouride.
  • Each membrane strip is divided into three sections 13a, 13b and 13c, each being coated with an antigene different from the antigenes on the other two sections, but corresponding to the antibodies to be detected.
  • Each membrane strip is thus adapted for simultaneous detection of 3 different antigenes from a blood sample from the same person.
  • the rib 11 is shaped as a tube so that the membrane battery may easily be fished out by means of a holder tool (not shown) comprising pins for sticking into the tubular rib.
  • the battery consists of a block 21 having a number of oblong grooved chambers or cells 22, being open at the top. The distance between the cells 22 is adjusted according to the distance between the flaps 12 and the membrane strips 13 with antigenes.
  • the block 21 is provided with a longitudinal recess 23, adapted for receiving the ribs 11 with flaps 12 and strips 13 of the membrane battery.
  • the block 21 is provided with another longi- tudinal recess 24, adapted for receiving a rib 31 on a holder for filter paper with blood samples, see fig. 11.
  • the said holder comprises a rib 31 with a number of flaps 32 with pieces of filter paper 33 being coated with blood samples.
  • Fig. 10 shows a section through a cell 22 with inserted holder for blood samples 31, 32, 33 and holder for membrane strips 11, 12, 13. Elution and incubation liquid 14 is introduced into the cells, and the culturing and aftertreatment are carried out under the same conditions as stated in example 1. After wash and colouring the membrane battery is removed and the test result read.
  • the examples are only illustrative and that the method can be varied in various ways, as will be readily appreciated by a person skilled in the art.
  • the method may be used not only for detecting antibodies in blood, but also for detecting antigenes in various liquids. In the latter case use may be made of strips with screened polyclonal antibodies or monoclonal antibodies.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (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)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Le procédé décrit, qui sert à détecter la présence d'anticorps dans le sang, utilise un échantillon de sang coagulé sur un support, tel qu'une goutte de sang sur un morceau de papier filtre, qui est introduit dans un récipient avec un liquide d'élution et d'incubation connu en soi et une bande de feuille recouverte d'antigènes correspondant au/aux type/types d'anticorps dont on désire détecter la présence. Après incubation et coloration, on effectue l'opération de détection pour déterminer une réaction positive ou négative. La présente invention se rapporte également à une bande de membrane ou à une batterie de membranes ainsi qu'à un liquide d'élution et d'incubation destinés à être utilisés dans ledit procédé et un kit de test comprenant l'équipement nécessaire pour effectuer l'analyse.
PCT/DK1989/000025 1988-02-11 1989-02-10 Procede servant a detecter la presence d'anticorps dans le sang et bande de membrane et batterie de membranes utilisees lors de la realisation dudit procede Ceased WO1989007763A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
BR898907241A BR8907241A (pt) 1988-02-11 1989-02-10 Processo para detectar anticorpos em uma amostra de sangue por tecnicas de imunoensaio,tira de membrana e bateria de membranas
FI903964A FI903964A0 (fi) 1988-02-11 1989-02-10 Foerfarande foer detektering av antikroppar i blod, och vid metoden anvaend membranremsa och membranuppsaettning.
NO90903535A NO903535L (no) 1988-02-11 1990-08-10 Fremgangsmaate for detektering av antistoffer i blod, membranstrimmel og membranbatteri for utfoerelse av fremgangsmaaten.
DK190390A DK190390A (da) 1988-02-11 1990-08-10 Fremgangsmaade til paavisning af antistoffer i blod, samt membranstrimmel og membranbatteri til anvendelse ved udoevelse af fremgangsmaaden
BG092796A BG92796A (bg) 1988-02-11 1990-09-06 Метод за откриване на антитела в кръвта,мембранна лента и мембранна батерия за провеждана на метода

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK704/88 1988-02-11
DK070488A DK70488D0 (da) 1988-02-11 1988-02-11 Fremgangsmaade til paavisning af antistoffer i blod, samt membran og test-saet til anvendelse ved udoevelse af fremgangsmaaden

Publications (1)

Publication Number Publication Date
WO1989007763A1 true WO1989007763A1 (fr) 1989-08-24

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ID=8096577

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Application Number Title Priority Date Filing Date
PCT/DK1989/000025 Ceased WO1989007763A1 (fr) 1988-02-11 1989-02-10 Procede servant a detecter la presence d'anticorps dans le sang et bande de membrane et batterie de membranes utilisees lors de la realisation dudit procede

Country Status (11)

Country Link
EP (1) EP0398989A1 (fr)
CN (1) CN1037036A (fr)
AU (1) AU3188089A (fr)
BG (1) BG92796A (fr)
BR (1) BR8907241A (fr)
DK (1) DK70488D0 (fr)
FI (1) FI903964A0 (fr)
IL (1) IL89225A0 (fr)
MC (1) MC2100A1 (fr)
OA (1) OA09627A (fr)
WO (1) WO1989007763A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5244788A (en) * 1992-07-01 1993-09-14 Hubscher Thomas T Method and apparatus for performing determinations of immune rectants in biological fluids
EP0574243A3 (en) * 1992-06-10 1994-05-25 Labsystems Oy Analytical transfer device
US5733775A (en) * 1995-02-27 1998-03-31 Slt Labinstruments Gesellschaft M.B.H. Temperature control device

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106501528A (zh) * 2016-10-27 2017-03-15 杭州量康科技有限公司 基于滤纸干血斑检测ToRCH10项抗体的方法
CN106990233B (zh) * 2017-05-27 2019-05-07 于宁 一种免疫检测凸泡板

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2099578A (en) * 1981-04-29 1982-12-08 Ciba Geigy Ag Supported reagents and kits for immunological analysis
EP0174247A2 (fr) * 1984-08-23 1986-03-12 Bernard Guerin Bandelette d'analyse immunologique et procédé pour sa fabrication
EP0217583A1 (fr) * 1985-09-23 1987-04-08 VXR, Inc. Extraction simultanée d'un ligand d'un échantillon et capture par un antiligand à cet effet, dans des essais ligands/antiligands
WO1987003965A1 (fr) * 1985-12-18 1987-07-02 Celldynamics Ag Bandelettes multi-indicatrices pour analyse immunologique, leur utilisation et leur fabrication

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2099578A (en) * 1981-04-29 1982-12-08 Ciba Geigy Ag Supported reagents and kits for immunological analysis
EP0174247A2 (fr) * 1984-08-23 1986-03-12 Bernard Guerin Bandelette d'analyse immunologique et procédé pour sa fabrication
EP0217583A1 (fr) * 1985-09-23 1987-04-08 VXR, Inc. Extraction simultanée d'un ligand d'un échantillon et capture par un antiligand à cet effet, dans des essais ligands/antiligands
WO1987003965A1 (fr) * 1985-12-18 1987-07-02 Celldynamics Ag Bandelettes multi-indicatrices pour analyse immunologique, leur utilisation et leur fabrication

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Clin. Chem. Vol. 27, No. 8, (1981), pages 1421-1423, (K. MIYAI et al.) "Two-Site Immunoenzymometric Assay for Thyrotropin in Dried Blood Samples on Filter Paper", see page 1422, "Assay Procedures". *
Clin. Chem. Vol. 32, No. 11, (1986), pages 2079-2082, (D. TSAO et al.) "Two-Site Enzyme Immunoassay for Alpha-Fetoprotein in Dried-Blood Samples Collected on Filter Paper", see page 2080 "Assay Procedure". *
Electrophoresis, Vol. 7, 1986, pages 1-18 (U. BEISIEGEl) "Protein Blotting", see paragraph 7.2.5 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0574243A3 (en) * 1992-06-10 1994-05-25 Labsystems Oy Analytical transfer device
US5474742A (en) * 1992-06-10 1995-12-12 Labsystems Oy Apparatus for removing solid bodies from an array of reaction vessels
US5244788A (en) * 1992-07-01 1993-09-14 Hubscher Thomas T Method and apparatus for performing determinations of immune rectants in biological fluids
US5733775A (en) * 1995-02-27 1998-03-31 Slt Labinstruments Gesellschaft M.B.H. Temperature control device

Also Published As

Publication number Publication date
OA09627A (en) 1993-04-30
AU3188089A (en) 1989-09-06
FI903964A7 (fi) 1990-08-10
IL89225A0 (en) 1989-09-10
MC2100A1 (fr) 1991-02-15
FI903964A0 (fi) 1990-08-10
BG92796A (bg) 1993-12-24
CN1037036A (zh) 1989-11-08
DK70488D0 (da) 1988-02-11
EP0398989A1 (fr) 1990-11-28
BR8907241A (pt) 1991-03-05

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