EP0760099A1 - Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique - Google Patents

Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique

Info

Publication number
EP0760099A1
EP0760099A1 EP94917941A EP94917941A EP0760099A1 EP 0760099 A1 EP0760099 A1 EP 0760099A1 EP 94917941 A EP94917941 A EP 94917941A EP 94917941 A EP94917941 A EP 94917941A EP 0760099 A1 EP0760099 A1 EP 0760099A1
Authority
EP
European Patent Office
Prior art keywords
recombinant
bacterial enzyme
denatured
diluent
assay
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
EP94917941A
Other languages
German (de)
English (en)
Other versions
EP0760099A4 (fr
Inventor
Robert C. Schoen
Yuzo Inoue
Toshinori Takei
Satoshi Jomura
Susan E. Sweeney
Joseph S. Niedbalski
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.)
Abbott Japan Co Ltd
Abbott Laboratories
Original Assignee
Dainabot Co Ltd
Abbott Laboratories
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 Dainabot Co Ltd, Abbott Laboratories filed Critical Dainabot Co Ltd
Publication of EP0760099A1 publication Critical patent/EP0760099A1/fr
Publication of EP0760099A4 publication Critical patent/EP0760099A4/fr
Ceased legal-status Critical Current

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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • 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
    • G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
    • 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
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G01N33/56988—HIV or HTLV
    • 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
    • G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
    • G01N33/5767—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis non-A, non-B hepatitis

Definitions

  • the invention relates generally to a method for improving the specificity of immunoassays, and more particularly, relates to a method for adding denatured and/or purified bacterial enzyme to a specimen diluent to increase the specificity of immunoassays which utilize recombinant antigens expressed as fusion proteins with a bacterial enzyme.
  • Viral lysates or recombinant protein preparations which comprise the typical antigen solutions which are utilized on the solid support, also can contain other proteins. These other proteins can bind to the solid support, introducing the possibility of reactivity with antibodies in the patient's sample. Alternatively, a component in the patient's sample can bind to the solid support and interfere with the assay.
  • McFarlane et al. Lancst (1990) 335:754-57, reported a high prevalence of antibody to hepatitis C virus (HCV) in patients with autoimmune chronic active hepatitis (AI-CAH). They suggested that the serum of AI-CAH patients may contain a component that gives false-positive results in the assay. McFarlane et al.
  • the assay might have been non-specif ⁇ cally detecting IgG since they saw a correlation between IgG levels and OD values, or that factors contained in the patient sera were responsible for the results, such as an antibody against some other pathogen which cross-reacts with the antigen used in the assay, or a component which adheres to the solid phase and binds IgG.
  • factors contained in the patient sera were responsible for the results, such as an antibody against some other pathogen which cross-reacts with the antigen used in the assay, or a component which adheres to the solid phase and binds IgG.
  • some false positive results are due to cross-reactivity between the patient sera and the fusion protein used to express the recombinant antigen utilized in the assay.
  • the present invention provides an improved method for increasing specificity in immunoassays which comprises the steps of (a) mixing the specimen with a diluent comprising a denatured recombinant bacterial enzyme expressed as a fusion protein with said bacterial enzyme, and (b) contacting said diluted specimen with at least one recombinant antigen expressed as a fusion protein with said denatured bacterial enzyme.
  • the recombinant denatured bacterial enzyme comprises a recombinant protein, and can be either CKS or SOD.
  • the concentration of said recombinant denatured bacterial enzyme is from about 0.001 g/L diluent to about 1.0 g L diluent More preferably, the concentration of said recombinant denatured bacterial enzyme is from about 0.01 g L diluent to about 0.1 g/L diluent. Most preferred, the recombinant bacterial enzyme is at a concentration of about 100 ⁇ g/ml.
  • the method can detect numerous analytes, including antibody to hepatitis C virus (HCV) and antibody to human immunodeficiency virus (HTV). Denaturation can be accomplished by a variety of methods, including heat and urea. Further, purified, recombinant bacterial enzyme may be utilized.
  • the present invention also provides a diluent useful for detecting antibodies in a test sample when performing an assay which uses at least one recombinant antigen expressed as a fusion protein with a recombinant bacterial enzyme, which diluent comprises the recombinant bacterial enzyme which has been denatured.
  • concentration of said denatured recombinant bacterial enzyme is about 100 ⁇ g/ml.
  • Purified, recombinant bacterial enzyme may be utilized.
  • the present invention also provides a test kit for performing an immunoassay which comprises a container containing a denatured recombinant bacterial enzyme selected from the group consisting of CKS and SOD.
  • the present invention provides an improvement to methods for detecting antibodies in a specimen from an individual wherein a recombinant antigen employed in the method is expressed as a fusion protein by a vector and wherein the recombinant antigen is encoded with a gene of a bacterial enzyme such as CKS or SOD.
  • the improvement comprises the step of mixing the specimen with a diluent comprising denatured recombinant bacterial enzyme (such as, SOD or CKS), wherein said enzyme also may be purified by methods known in the art as discussed herein, including anion- exchange resins and cation-exchange resins.
  • Denaturation of proteins occurs when the tertiary bonds of proteins are broken, which results in partial or complete unfolding of the protein. Usually, however, care is taken not to denature the protein solutions used for assays, since such denaturation can change the properties of such proteins, adversely affecting the results of the assay. See, for example, N. W. Tietz, ed., Fundamentals of Clinical Chemistry. 2nd Edition, W. B. Saunders Company, Philadelphia (1976), page 270.
  • a false positive result is defined herein as one in which a sample is repeatably reactive in an ELISA, EIA or PHA but is not confirmed by alternative methods for the presence of analyte antibodies.
  • Alternative testing methods include synthetic peptide EIAs, antibody blocking procedures and recombinant immunoblot assays.
  • Applicants have observed that in certain instances, addition of recombinant or purified recombinant bacterial enzymes does not result in increased specificity in assays which employ recombinant proteins. Applicants have surprisingly discovered that denaturation of the recombinant bacterial enzyme results in increased specificity in assays. Also, applicants have discovered that at times it may be advantageous to utilize a purified, denatured, recombinant bacterial enzyme in a specimen diluent to increase the specificity of the assay.
  • Immunoassays can employ recombinant antigens which are expressed as a fusion protein by a vector in which the antigen is encoded with the superoxide dismutase (SOD) gene or the CKS (CTP:CMP-3-deoxy- -manno-octulosonate cytidylyl transferase or CMP-KDO synthetase) gene (see, U.S. Patent No. 5,124,255, issued June 23, 1992, which is incorporated herein by reference and U.S.
  • SOD superoxide dismutase
  • CKS CTP:CMP-3-deoxy- -manno-octulosonate cytidylyl transferase or CMP-KDO synthetase
  • the effect of this anti-CKS or anti-SOD is not removable as a false positive reaction unless the bacterial enzyme which is employed is denatured prior to its addition in the specimen diluent
  • the bacterial enzyme preferably recombinantly engineered, can be added to the diluent in concentrations ranging from about 0.001 g/L to about 1 g/L, more preferably from about 0.005 g/L to about 0.5 g/L, and most preferably from about 0.01 g/L to about 0.1 g/L.
  • the bacterial enzyme can be added as a purified or partially purified protein from a bacterial extract. However, it has been discovered that the bacterial enzyme must be denatured prior to addition and use in the diluent.
  • Denaturation can be effected by methods known in the art, including heating at approximately 50°C to 60°C for approximately 20 to 60 minutes, and treatment of the bacterial enzyme with chemicals, such as, 8M urea, guanidine, and some organic solvents.
  • chemicals such as, 8M urea, guanidine, and some organic solvents.
  • a "diluent” is defined herein as an aqueous solution of buffers) and salt(s) as well understood in the art and illustrated infra.
  • a preferred buffer is Trisftydroxymethyljaminomethane, commercially available under the trade designation Tris from Sigma Chemical Co., St.
  • suitable buffers include, but are not limited to, buffers such as HEPES (N-[2-hydroxye l]piperazme-N'-[2-ethanesulfonic acid]), PIPES (piperazine-N,N-bis[2-ethanesulfonic acid]), CAPS (3-[cyclohexylamino]-l- propanesulfonic acid) and MOPES (3-[N-morpholino]propanesulfonic acid).
  • Suitable salts include sodium chloride (NaQ) and salts such as phosphate salts and sulfate salts.
  • animal serum proteins such as bovine serum, bovine serum albumin (BSA), fetal calf serum and goat serum can be added in concentrations ranging from about 0.5% v v to about 50% v v.
  • Biological detergents such as polyoxyethylenesorbitan, commercially available as Tween® 20, polyoxyethylene ether, commercially available as Triton® X-100, Nonidet P-40 (an octylphenol-ethylene oxide condensate), sodium dodecyl sulfate (SDS) or N-lauroylsarcosine (N-dodecanoyl-N-methylglycine) can be added in concentrations ranging from about 0.01% v v to about 5% v/v.
  • polyoxyethylenesorbitan commercially available as Tween® 20
  • polyoxyethylene ether commercially available as Triton® X-100
  • Nonidet P-40 an octylphenol-ethylene oxide condensate
  • SDS sodium dodecyl sulfate
  • N-lauroylsarcosine N-dodecanoyl-N-methylglycine
  • Chelators such as ethylenediaminetetraacetic acid (EDTA) and ethylene glycol-bis( ⁇ -aminoethylether) N .KN'.N'-tetraacet-c acid (EGTA) can be added in concentrations ranging from about 2 mM to about 20 mM.
  • EDTA ethylenediaminetetraacetic acid
  • EGTA ethylene glycol-bis( ⁇ -aminoethylether) N .KN'.N'-tetraacet-c acid
  • a lymphocyte lysate solution for example, human T-lymphocyte solution or a host cell lysate solution such as an E. coli lysate solution, can be added in concentrations typically ranging from about 0.01% v/v to about 10% v v.
  • Preservatives such as sodium azide can also be added.
  • the specimen diluent can be used to dilute test specimens in various assay formats. It is contemplated that the diluent reagent employed for the assay can be provided in the form of a kit with one or more containers such as vials or bottles, with each container containing a separate reagent such as a diluent, a monoclonal antibody or combination thereof, or a recombinant protein employed in the assay.
  • Solid phases which can be used in the assay formats are known to those in the art and include the walls of wells of a reaction tray, test tubes, polystyrene beads, magnetic beads, nitrocellulose strips, membranes, microparticles such as latex particles, and others.
  • the “solid phase” is not critical and can be selected by one skilled in the art Thus, latex particles, microparticles, magnetic or non ⁇ magnetic beads, membranes, plastic tubes, walls of microtiter wells, glass or silicon chips and red blood cells are all suitable examples. Suitable methods for immobilizing peptides on solid phases include ionic, hydrophobic, covalent interactions and the like.
  • the present invention provides a diluent, test kit and method for increasing specificity in immunoassays which utilize specific binding members.
  • a "specific binding member,” as used herein, is a member of a specific binding pair. That is, two different molecules where one of the molecules through chemical or physical means specifically binds to the second molecule. Therefore, in addition to antigen and antibody specific binding pairs of common immunoassays, other specific binding pairs can include biotin and avidin, carbohydrates and lectins, complementary nucleotide sequences, effector and receptor molecules, cofactors and enzymes, enzyme inhibitors and enzymes, and the like. Furthermore, specific binding pairs can include members that are analogs of the original specific binding members, for example, an analyte- analog.
  • Immunoreactive specific binding members include antigens, antigen fragments, antibodies and antibody fragments, both monoclonal and polyclonal, and complexes thereof, including those formed by recombinant DNA molecules.
  • hapten refers to a partial antigen or non-protein binding member which is capable of binding to an antibody, but which is not capable of eliciting antibody formation unless coupled to a carrier protein.
  • analyte is the substance to be detected which may be present in the test sample.
  • the analyte can be any substance for which there exists a naturally occurring specific binding member (such as, an antibody), or for which a specific binding member can be prepared.
  • an analyte is a substance that can bind to one or more specific binding members in an assay.
  • “Analyte” also includes any antigenic substances, haptens, antibodies, and combinations thereof.
  • the analyte can be detected by means of naturally occurring specific binding partners (pairs) such as the use of intrinsic factor protein as a member of a specific binding pair for the determination of Vitamin B 12, the use of folate-binding protein to determine folic acid, or the use of a lectin as a member of a specific binding pair for the determination of a carbohydrate.
  • the analyte can include a protein, a peptide, an amino acid, a hormone, a steroid, a vitamin, a drug including those administered for therapeutic purposes as well as those administered for illicit purposes, a bacterium, a virus, and metabolites of or antibodies to any of the above substances.
  • Viruses which can be tested include hepatitis-causing viruses (for example , hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis delta, and hepatitis E virus), human immunodeficiency viruses (such as HTV-1, HTV-2), the HTLV-I and HTLV-II viruses, and the like.
  • hepatitis-causing viruses for example , hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis delta, and hepatitis E virus
  • human immunodeficiency viruses such as HTV-1, HTV-2
  • the HTLV-I and HTLV-II viruses and the like.
  • the “indicator reagents” may be used in the various assay formats discussed herein.
  • the “indicator reagent” “comprises a “signal generating compound” (label) which is capable of generating a measurable signal detectable by external means conjugated (attached) to a specific binding member for the analyte.
  • Specific binding member as used herein means a member of a specific binding pair. That is, two different molecules where one of the molecules through chemical or physical means specifically binds to the second molecule.
  • the indicator reagent also can be a member of any specific binding pair, including either hapten-anti-hapten systems such as biotin or anti-biotin, avidin or biotin, a carbohydrate or a lectin, a complementary nucleotide sequence, an effector or a receptor molecule, an enzyme cofactor and an enzyme, an enzyme inhibitor or an enzyme, and the like.
  • hapten-anti-hapten systems such as biotin or anti-biotin, avidin or biotin, a carbohydrate or a lectin, a complementary nucleotide sequence, an effector or a receptor molecule, an enzyme cofactor and an enzyme, an enzyme inhibitor or an enzyme, and the like.
  • An immunoreactive specific binding member can be an antibody, an antigen, or an antibody/antigen complex that is capable of binding either to the analyte as in a sandwich assay, to the capture reagent as in a competitive assay, or to the ancillary specific binding member as in an indirect assay.
  • labels include chromogens, catalysts such as enzymes, luminescent compounds such as fluorescein and rhodamine, chemiluminescent compounds, radioactive elements, and direct visual labels.
  • enzymes include alkaline phosphatase, horseradish peroxidase, beta-galactosidase, and the like. The selection of a particular label is not critical, but it will be capable of producing a signal either by itself or in conjunction with one or more additional substances.
  • test sample includes biological samples which can be tested by the methods of the present invention described herein and include human and animal body fluids such as whole blood, serum, plasma, cerebrospinal fluid, urine, lymph fluids, and various external secretions of the respiratory, intestinal and genitourinary tracts, tears, saliva, milk, white blood cells, myelomas and the like, biological fluids such as cell culture supernatants, fixed tissue specimens and fixed cell specimens. Any substance which can be diluted and tested with recombinant proteins and assay formats described in the present invention are contemplated to be within the scope of the present invention. It is contemplated that whatever assay format is chosen will utilize a diluted test sample.
  • haptens are known in the art It is contemplated that haptens also can be used in assays employing fusion proteins in order to enhance performance of the assay.
  • haptens also can be used in assays employing fusion proteins in order to enhance performance of the assay.
  • the following examples are meant to illustrate, but not to limit, the spirit and scope of the invention.
  • DEAE ion-exchange resin or a QAE ion-exchange resin was filled up in the column, e.g., DEAE-5PW (available from Toso), Resource-Q (available from Pharmacia), was equilibrated with the 10 mM phosphate buffer containing 8M urea (pH 7.5).
  • CKS-rp 36 antigen in 8M urea solution was passed over the above anion-exchange resin column.
  • CKS-rp 36 antigen was passed by this step at a flow rate of lml/min.
  • the resultant CKS-rp 36 antigen was used in a passive hemagglutation assay (PHA) as an antigen, as follows.
  • PHA passive hemagglutation assay
  • CM ion-exchange resin or SP ion-exchange resin was filled up in columns, e.g., CM-3WS (available from Toso), Resource-S (available from Pharmacia) and equilibrated with the 10 mM phosphate buffer containing 8M urea (pH 7.5).
  • CKS-rp 36 antigen was passed over the above cation exchange resin column. The column then was extensively washed, and the CKS-rp 36 antigen was eluted by step elusion with salt at a flow rate of 1 ml/min.
  • the denatured CKS fusion protein then was coated onto human red blood cells type O, and used as an antigen in the PHA assay described hereinabove.
  • Three thousand (3000) volunteer serum donors were tested using the PHA kit described hereinabove and wherein the test serum was diluted at various concentrations. The results obtained were as summarized in Table 2.
  • Example 2 Preparation of HTV-Related Antibody Assay Kit Human red blood cells type O first were washed with physiological sale, pH 7.0, and then incubated with a solution containing DEAE-CM treated HTV CKS-rp 36 antigen. The cells were incubated in the antigen solution for two hours at room temperature and washed in phosphate buffered saline. The antigen coated cell solution consisted of 1.0% (v/v) of the antigen coated cells in physiological saline solution. E . coli XL-1 lysates, E. coli JM 103 lysates and E.
  • Example 4 Specimen Diluent
  • the sample diluent comprising 10% (v v) bovine serum and 20% (v/v) goat serum in 20 mM Tris phosphate buffer containing 0.15% (v/v) Triton X-100, 1% (w/v) BSA, and 500 ⁇ g ml or less recombinant CKS bacterial enzyme (prepared as described in Example 3) was prepared, wherein the recombinant CKS bacterial enzyme was denatured at 50°C for approximately 30 minutes before adding to the diluent
  • the Abbott 2nd generation HTV assay (available from Abbott Laboratories, Abbott Park, IL) was used in the experiment Specimens were run in duplicate, with controls being run five times each. The comparison was made between the diluent available as the standard assay reagent and a diluent which contained CKS recombinant bacterial enzyme (denatured at 50°C for 30 minutes), as described in Example 4. Assay protocol followed was as recommended by the manufacturer of the assay, with readings obtained reported as optical density readings (O.D.). The results are summarized in the following Table 5. TABLE 5
  • Example 6 Specificity Panel Testing for HTV 1/HTV 2 Test Pack Assay
  • HTVl/H ⁇ V2 Assay Previous data had shown that the addition of CKS lysate to the HTV1/HTV2 Test Pack Assay (available from Abbott Laboratories, Abbott Park, IL) did not increase specificity in the HTV assay by reducing the number of false positive reactions believed to be occurring. Consequently, experiments were designed to determine whether denaturation of the recombinant CKS bacterial enzyme would improve specificity.
  • a specificity panel consisting of the same 10 members described in Example 7 was selected for testing, along with positive and negative controls.
  • the Abbott HTV 1 HTV 2 Test Pack assay (available from Abbott Laboratories, Abbott Park, IL) was used in the experiment Specimens including positive and negative controls were run in duplicate, and various concentrations of heat treated CKS recombinant bacterial enzyme were tested (12 ⁇ g/ml, 25 ⁇ g/ml, 50 ⁇ g/ml, 75 ⁇ g/ml and 100 ⁇ g/ml). All heat treatment to denature the CKS bacterial enzyme was performed at 50°C for 30 minutes. Assay protocol followed was as recommended by the manufacturer of the assay, with readings obtained reported as positive, negative or positive/negative.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • Virology (AREA)
  • Communicable Diseases (AREA)
  • AIDS & HIV (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

L'invention concerne un procédé amélioré de détection d'anticorps. Ce procédé comprend les étapes suivantes: a) mélange du spécimen avec un diluant comprenant peroxyde dismutase et b) mise en contact du spécimen dilué avec au moins un antigène recombinant exprimé en tant que protéine de fusion avec peroxyde dismutase.
EP94917941A 1994-05-09 1994-05-09 Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique Ceased EP0760099A4 (fr)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US1994/005152 WO1995030902A1 (fr) 1994-05-09 1994-05-09 Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique

Publications (2)

Publication Number Publication Date
EP0760099A1 true EP0760099A1 (fr) 1997-03-05
EP0760099A4 EP0760099A4 (fr) 1998-12-09

Family

ID=22242549

Family Applications (1)

Application Number Title Priority Date Filing Date
EP94917941A Ceased EP0760099A4 (fr) 1994-05-09 1994-05-09 Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique

Country Status (4)

Country Link
EP (1) EP0760099A4 (fr)
JP (1) JPH10504098A (fr)
AU (1) AU6946394A (fr)
WO (1) WO1995030902A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2293677C (fr) * 1998-04-14 2009-01-27 Otsuka Pharmaceutical Co., Ltd. Procedes de detection d'anticorps et dispositif de detection d'anticorps

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0388232A1 (fr) * 1989-03-17 1990-09-19 Chiron Corporation Diagnostics et vaccins de NANBV
WO1992013275A1 (fr) * 1991-01-25 1992-08-06 Abbott Laboratories Utilisation de superoxyde-dismutase dans un diluant d'echantillon
WO1994000594A1 (fr) * 1992-06-23 1994-01-06 Abbott Laboratories Procedes d'utilisation des proteines de fusion cks

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5124255A (en) * 1988-03-11 1992-06-23 Abbott Laboratories CKS method of protein synthesis
JP3307637B2 (ja) * 1991-03-06 2002-07-24 オーソ ダイアグノスティック システムズ インコーポレイテッド 酵素結合抗体免疫吸着アッセイを改善するために変性した伝播体タンパク質

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0388232A1 (fr) * 1989-03-17 1990-09-19 Chiron Corporation Diagnostics et vaccins de NANBV
WO1992013275A1 (fr) * 1991-01-25 1992-08-06 Abbott Laboratories Utilisation de superoxyde-dismutase dans un diluant d'echantillon
WO1994000594A1 (fr) * 1992-06-23 1994-01-06 Abbott Laboratories Procedes d'utilisation des proteines de fusion cks

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO9530902A1 *

Also Published As

Publication number Publication date
AU6946394A (en) 1995-11-29
WO1995030902A1 (fr) 1995-11-16
EP0760099A4 (fr) 1998-12-09
JPH10504098A (ja) 1998-04-14

Similar Documents

Publication Publication Date Title
US4588680A (en) Assay for viruses
CN101419238B (zh) 丙型肝炎病毒核心抗原化学发光酶联免疫检测试剂盒
AU674353B2 (en) Base dissociation assay
Chau et al. Detection of IgA class antibody to hepatitis E virus in serum samples from patients with hepatitis E virus infection
WO1993021346A1 (fr) Dosage pour la detection de l'antigene du vih et de l'anticorps du vih
JPH04233461A (ja) 同時アッセイ法
US6511812B1 (en) Method and test kit for use in improving immunoassay specificity
EP1083428A2 (fr) Méthode et réactif pour la détection ou détermination des antigènes du noyau de HCV
KR100500793B1 (ko) C형간염바이러스감염증진단약
vd Waart et al. Enzyme‐immunoassay in the diagnosis of hepatitis with emphasis on the detection of “e” antigen (HBeAg)
US4814269A (en) Diagnostic testing for antibodies against microorganisms
WO1995030902A1 (fr) Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique
Ivanov et al. Enzyme-linked immunosorbent assay for detection of arenaviruses
AU658395B2 (en) Use of superoxide dismutase in specimen diluent
CA2005204C (fr) Immuno-essai en phase solide avec conjugue lyophilise
US5149623A (en) Rapid, easy, and economical screening test for antibodies to human immunodeficiency virus
Garland et al. A solid-phase enzyme-linked immunosorbent assay (ELISA) for detection of antibody to rubella virus
JP4433642B2 (ja) 非特異反応を抑制したイムノアッセイ法
KR101561893B1 (ko) 잠재 헤파타이티스 ｃ의 검출을 위한 개선된 분석 방법, 그 사용 및 대응하는 진단 키트
WO1995002186A1 (fr) Nouveau dosage diagnostique destine a la detection de la syphilis
RU2056057C1 (ru) Способ получения иммобилизованного иммуноглобулина на твердой фазе
AU664179B2 (en) Immunoassay for detecting HCV IgM antibody
RU2084527C1 (ru) Набор для определения антител к вирусу гепатита c человека (варианты)
CA2189748A1 (fr) Procede et reactifs permettant d'ameliorer la specificite d'un dosage immunologique
CA1338528C (fr) Essai de depistage rapide, facile et peu couteux pour les anticorps du virus de l'immunodeficience humaine

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19961018

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE ES FR GB IT LI NL

A4 Supplementary search report drawn up and despatched

Effective date: 19981022

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH DE ES FR GB IT LI NL

17Q First examination report despatched

Effective date: 20000307

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED

18R Application refused

Effective date: 20040219