WO2001003721A1 - Dosages ameliores de detection d'infection au virus de l'immunodeficience humaine - Google Patents

Dosages ameliores de detection d'infection au virus de l'immunodeficience humaine Download PDF

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Publication number
WO2001003721A1
WO2001003721A1 PCT/US2000/019017 US0019017W WO0103721A1 WO 2001003721 A1 WO2001003721 A1 WO 2001003721A1 US 0019017 W US0019017 W US 0019017W WO 0103721 A1 WO0103721 A1 WO 0103721A1
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Prior art keywords
hiv
antigen
avidin
receptor
epitope
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PCT/US2000/019017
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English (en)
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Gregory Cullen
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Merlin Biomedical & Pharmaceutical, Ltd.
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Priority to AU62111/00A priority Critical patent/AU6211100A/en
Publication of WO2001003721A1 publication Critical patent/WO2001003721A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16211—Human Immunodeficiency Virus, HIV concerning HIV gagpol
    • C12N2740/16222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention generally relates to assays to determine if an individual has been infected by the human immunodeficiency virus (HIV). More particularly, the present invention relates to modified HIV antigens and corresponding receptors which may be incorporated into current HIV assay systems to increase the sensitivity of the assays.
  • HIV human immunodeficiency virus
  • HIV is the causative agent of Acquired Immunodeficiency Syndrome, commonly referred to by its acronym AIDS.
  • HIV 1 and HIV 2 There are two major strains of virus designated HIV 1 and HIV 2.
  • HIV 1 and HIV 2 When an individual is infected with either strain of HIV, that person's immune system will develop antibodies which recognize and bind to certain portions of the virus. These portions are known as antigens or epitopes.
  • antigens or epitopes The presence of anti-HIV antibodies in blood is therefore a reliable indicator of HIV infection.
  • the most common antibodies detected in individuals infected with HIV are directed to the core antigen p24; the polymerase antigen p51; and the envelope antigens gp120, gp41 and gp36. Almost all individuals with either HIV 1 or HIV 2 infection will have anti-HIV antibodies to one or more of these antigens.
  • the standard method of screening for HIV infection is based on an enzyme-immunoassay (EIA) procedure to detect anti-HIV antibodies in a blood sample.
  • EIA enzyme-immunoassay
  • HIV antigens are first "fixed" directly to the interior wall of a microwell either by adsorption or by chemical reaction. The blood sample to be tested is then added.
  • any anti-HIV antibodies are present they will bind to the fixed HIV antigens. Subsequently, an enzyme reagent is typically added to indicate if any anti-HIV antibodies are bound to the HIV antigens.
  • immunochromatographic assays have been developed.
  • a blood sample is placed on a specially prepared membrane strip.
  • the strip has adsorbed colloidal gold particles bearing HIV antigen nearer to the end of the strip where the blood sample is placed, and further down has fixed HIV antigen bound to the strip.
  • the constituents of the blood sample migrate along the strip by wickmg or capillary action. Any HIV antibodies in the blood sample will bind to the HIV antigens on the colloidal gold particles.
  • the HIV antibody bound gold particles then continue to migrate along the membrane strip until they contact the band of fixed HIV antigens on the strip.
  • Some of the HIV antibody bound to the gold particles have free binding sites and these will bind to the fixed HIV antigens on the membrane thus immobilizing the gold particles to form a visible band.
  • a second control band of reagents that will bind the excess HIV bound gold particles at the distal end of the strip serves to demonstrate that the test is performing correctly.
  • the EIA and immunochromatographic assay have not been optimized to increase the sensitivity and accuracy of the assays
  • Current methods of attaching the HIV antigens to the solid supports (i.e. microwelis or membranes, etc.) using adsorption or chemical means often result in the HIV antigens being inappropriately attached, such as. — the HIV molecule may be folded with the HIV epitope buried in the interior; - the HIV epitope may be attached with the inactive end exposed; or
  • the HIV epitope may be buried under protein molecules.
  • the present invention provides reagents which may be incorporated into conventional HIV assays, thereby significantly increasing the sensitivity of the assay. Consequently, HIV assays modified in accordance with the present invention are more accurate. Furthermore, because the present invention optimizes the use of HIV antigens in an assay, less antigen is required, and the assay may be manufactured at a lower cost.
  • a method of detecting the presence or absence of anti-HIV antibodies in a sample of human blood comprises providing a substrate bearing a plurality of exposed avidin receptor molecules.
  • the avidin receptor molecules are then bound to a plurality of biotinylated HIV antigens to form a plurality of receptor-HIV antigen complexes.
  • the complexes are then exposed to human blood under conditions which permit anti-HIV antibodies to bind to the HIV antigen portion of the complex.
  • the presence or absence of bound anti-HIV antibodies may then be detected with conventional means such as enzymeimmu ⁇ oassay or immunochromatographic assay, as for example horseradish peroxidase assay.
  • an anti-HIV antibody binding complex consists of a plurality of avidin receptors fixed to a substrate. Bound to the avidin receptors are at least one type of HIV antigen modified by addition of a biotm group. In a preferred embodiment, the biotin group is added to the N-termi ⁇ al ammo acid of the HIV antigen.
  • the HIV antigen may be selected from the group consisting of p24, p51, gp 120, gp 41, gp 36, and combinations thereof.
  • the substrate may be any substrate used in conventional HIV assays to bind to HIV antigens, such as microwells or colloidal gold particles.
  • a biotinylated HIV antigen in another aspect of the present invention, there is provided a biotinylated HIV antigen.
  • the biotinylated HIV antigen is capable of being bound by an avidin receptor molecule such that an epitope of the antigen is exposed for binding to an anti-HIV antibody.
  • the antigen may be p24 of HIV 1, and the epitope is formed from ammo acid residues 201 217 of p 24.
  • the antigen may be gp 41 of HIV 1, where the epitope is formed from ammo acid residues 584-618 of gp 41.
  • the antigen may be gp 36 of HIV 2, where the epitope is formed from ammo acid residues 574-603 of gp 36.
  • Figures 1A-1 C are a schematic diagram of an anti-HIV antibody assay (enzymeimmu ⁇ oassay) incorporating the present invention.
  • FIG. 2 is a schematic diagram of a membrane assay system incorporating the present invention. Detailed Description of the Preferred Embodiment
  • the present invention comprises HIV antigens that can be firmly attached to a variety of solid supports in a precise manner.
  • the HIV antigens are attached through a process which controls their orientation and positioning, advantageously increasing the availability of epitopes for anti-HIV antibody binding in comparison to conventional attachment methods. Consequently, HIV assays which incorporate the teachings of the present invention are more sensitive for a given concentration of HIV antigen.
  • the present invention is used to modify the core antigen p24; the polymerase antigen p51; and the envelope antigens gp120, gp41 and gp36, as anti-HIV antibodies to these antigens are commonly found in those infected by HIV.
  • the present invention can be applied to a variety of HIV antigens.
  • the present invention accomplishes these benefits by the use of a liga ⁇ d-receptor system in which the HIV antigens are conjugated to a ligand which is specific for a receptor, which may then be fixed to a separate solid support or to other elements of the selected assay.
  • the ligand is conjugated to the HIV antigen on a portion of the antigen which is sufficiently far from the epitope as not to interfere with anti-HIV antibody binding.
  • the ligand group on the HIV protein is then allowed to react with the binding site on the receptor molecule.
  • the ligand and receptor are chosen such that the association binding constant of the ligand-receptor system is extremely high and essentially irreversible under assay conditions. Consequently, the HIV antigen is firmly bound and oriented so that the HIV antigen epitope is freely accessible to be bound by HIV antibodies.
  • the present invention is illustrated in schematic form.
  • the present invention may consist of a solid support 10.
  • a receptor molecule 20 is attached to solid support 10.
  • Receptor molecule 20 may be attached to solid support 10 in any manner which does not interfere with the ability of receptor molecule 20 to bind the ligand for which it is specific, as discussed in more detail below.
  • receptor molecule 20 may be bound to solid support 10 by chemical reaction producing a covalent bond, by adsorbtion, or by other means known to those of skill in the art.
  • solid support 10 is a microwell, of the type conventionally used to perform immunoassa ⁇ s. It should be appreciated by those of skill in the art, however, that solid support 10 can take a variety of different shapes and forms, and for purposes of the present invention, solid support 10 can be any solid support presently used to conduct HIV assays, such as wells and membranes.
  • a modified HIV antigen 30 has been introduced to solid support 10.
  • Modified HIV antigen 30 has an epitope
  • Modified HIV antigen 30 differs from HIV antigen normally found in blood by virtue of a ligand 32 which has been conjugated to the antigen.
  • Ligand 32 is specifically chosen to correlate with receptor molecule 20, such that ligand 32 will strongly bind to receptor 20.
  • modified HIV antigen 30 is bound to receptor molecule 20, with epitope 35 clearly exposed for binding to a ⁇ ti HIV antibodies.
  • Modified antigen 30 and receptor molecule 20 therefore form a complex.
  • the affinity of receptor 20 for antigen ligand 32 is high enough that the complex will not disassociate under assay conditions.
  • the disassociation constant (K dl of the receptor for the ligand is at least about 10 ' 5 or less.
  • FIG. 1 B there is shown a subsequent step in an HIV assay which incorporates the present invention.
  • solid support 10 and its receptor -antigen complexes have been exposed to blood containing anti-HIV antibodies.
  • anti-HIV antibody 50 binds to epitope 35 of the modified HIV antigen 30 via region 55.
  • FIG 1 C a later step in the assay is depicted, whereby an indicator antibody 60 has been added to solid support 10.
  • Indicator antibody 60 recognizes epitope 57 on anti-HIV antibody 50, and through region 62 binds to epitope 57.
  • Indicator antibody 60 possesses indicator means, such as an enzyme 65, which reacts with substrate 70 to produce colored molecule 75 which may then be viewed by the clinician performing the assay, and which indicates the presence of anti-HIV antibodies in the blood being tested.
  • receptor molecules 20 are avidin receptors.
  • the avidin molecule is well known to those of skill in the art.
  • Avidin may be obtained from a variety of sources.
  • avidin receptor can be isolated from egg whites, as strepavidin from streptomyces avidinn, or in recombinant form from various manufacturers.
  • strepavidin is preferred over other types of avidin, because it does not have carbohydrates which have been noted to result in non-specific binding.
  • Avidin receptors found suitable to practice the present invention include product ⁇ os. A9390 and A9275, available from Sigma BioSciences.
  • Avidin receptor molecules can be firmly attached to the solid supports by a variety of methods, including chemical bonding and adsorption, as known to those of skill in the art.
  • an excess of avidin receptor molecules may be used without interfering with the present invention. Any loss of binding activity of the receptor molecules due to chemical inactivatio ⁇ or poor orientation will not be significant because there will be sufficient receptor molecules remaining to bind the ligand labeled HIV antigens in their proper orientation. Also, because less HIV antigens are used there is less background and increased assay sensitivity.
  • the HIV antigens are modified by the addition of a biotin group which can bind to the avidin.
  • the biotin is covendingl ⁇ linked to the N-terminal ammo acid of the antigen peptide to be modified.
  • the preferred HIV antigens for the present invention are those which are most recognizable by antibodies in the blood of HIV infected individuals. As noted above, these antigens are found on the following HIV 1 and HIV 2 proteins: core antigen p24; the polymerase antigen p51 ; and the envelope antigens gp120, gp41 and gp36.
  • any assay incorporating the modified HIV antigens of the present invention includes a mixture of the commonly recognized antigens of HIV 1 and HIV 2, so that one assay may be used to screen for infection by either strain of the virus.
  • a purified mixture of HIV 1 p24 and gp41 and HIV 2 gp 36 may be used.
  • Alternate embodiments may include the p51 antigen.
  • Other mixtures may be used as well, provided that at least one commonly recognizable antigen from both HIV 1 and HIV 2 is included in the mixture.
  • mixtures of antigens it is preferred that approximately equal amounts of each antigen be used in the assay.
  • HIV antigens suitable to practice the present invention are those which are commercially available and which are recognized by anti-HIV antibodies.
  • the novel HIV antigens may be made by protein synthesis or recombi ⁇ a ⁇ t DNA techniques.
  • the preferred p24 antigen is a 24,000 MW group specific antigen from the capsid of the HIV-1 virus.
  • the p24 antigen is composed of ammo acids 1-258 of p24 and is expressed in a Baculovirus system. The protein is purified by column chromatography.
  • a recombinant gp41 antigen may also be used.
  • the gp41 antigen is a tra ⁇ smembrane protein specific for the HIV-1 virus, and comprises ammo acids 1 -5, 31 -168 and 194-241 of gp41. Ammo acid residues 6-30 are replaced by a single senne residue, and ammo acids 169-193 are deleted.
  • the recombinant gp41 is expressed in an E. coll system, and is purified by preparative eiectrophoresis.
  • the gp36 antigen is preferably the transmembra ⁇ e protein specific for the HIV-2 virus.
  • a recombinant gp36 may be used.
  • the recombinant protein may be composed of am o acids 43-148 of gp36.
  • the recombinant protein is expressed in an E. co system, and purified with an amylose column.
  • Biotinylatio ⁇ is based on the use of biotinamidocaproate-N-hydroxysulfosuccinimide ester. The reaction proceeds at pH 7.2 under mild conditions and proteins may be labeled with 2-5 moles of biotin per mole of protein. The biotinylated proteins may be separated from unreacted reagents by gel-filtration. Biot ylation of the antigens may be performed by suitable processes know to those of skill in the art, provided that the process does not significantly degrade the ability of an anti-HIV antibody to bind to the epitope of the HIV antigen which is being biotinylated. For example, biotmylation may be performed using commercially available reagents, such as those sold by Sigma BioSciences. One suitable process for biotmylation of the N-terminal ammo acid of HIV antigens is as follows:
  • reaction vessel may be washed with a few drops of PBS and added to the top of the column.
  • Protein containing fractions up to 66% of a column volume may be combined. For the column supplied, collecting 0.5 ml fractions, this would be fraction number 12. If fractions past this point are combined, they may contain free biotinylation reagent.
  • the modified HIV antigens may be formed by peptide synthesis.
  • This method of attaching the ligand group to the HIV epitope is to synthesize the ligand group at the terminal end of the synthetic HIV peptide.
  • the method of peptide synthesis utilizes a totally automated chemical synthesis process to build up the molecule one amino acid at a time.
  • the sequence of ami ⁇ o acids in the peptide depends on the sequence of nucieotide bases in the DNA chain. A particular sequence of three bases (or codon) is the code to form one particular ami ⁇ o acid, and the sequence of amino acids make up the peptide of interest. It is therefore possible to manufacture a HIV epitope molecule so that the ligand group is at the terminal end of the recombinant fusion protein and will not interfere with the binding of antibody to the epitope in the assay.
  • the first step involves preparing a custom DNA sequence that will code for the HIV epitope and include a ligand group at the terminal end of the recombinant fusion protein.
  • the specific DNA molecule is prepared using DNA polymerase and spliced to certain promoter genes to enhance yield.
  • the DNA+ promoter genes is then inserted into the genome of a plasmid vector.
  • the plasmid then "infects" a bacterium and inserts the HIV+ promoter genes into the bacterial DNA which become programmed to produce large quantities of HIV proteins.
  • the bacteria are allowed to multiply in fermentation tanks and the HIV proteins are collected and purified.
  • the first step is to attach the receptor (avidin) to the walls of the microwells.
  • the next step is to attach the biotinylated ligand (the novel HIV epitopes) to the fixed avidin.
  • Avidin is diluted in coating buffer (carbonate-bicarbonate pH 9.6), and 0.2 ml is pipetted into each microweli (Dynatech #011 -010-6302) and allowed to absorb at room temperature for several hours. The microwells are then washed three times with phosphate buffered saline and Tween solution (PBS-Tween) to remove unbonded material.
  • coating buffer carbonate-bicarbonate pH 9.6
  • PBS-Tween solution phosphate buffered saline and Tween solution
  • the patient's serum or plasma is diluted in PBS-Tween (usually 1 :100) and 0.2 mi is pipetted into the prepared microweli, and allowed to react at RT (usually 1 hr is sufficient).
  • the microwells are then washed 3 times with PBS-Tween.
  • 0.2 ml of enzyme labeled antibody conjugate (anti-human IgG or IgM antibody labeled with horse radish peroxidase e ⁇ zyme-' ⁇ RPO-conjugate") is added and allowed to react within any bound antibody (usually 1 hr at
  • microwells are washed 3 times with PBS-Tween and 0.2 ml of the substrate-chromogen solution (tetramethylbenzidine "TMB") is added and allowed to react. Positive results usually give a blue color change within 15 minutes and these can be measured in a spectrometer (microplate reader) at 655 nm (or H 2 S0 4 acid can be added which changes the blue color to yellow and can then be read at 450 nm).
  • TMB tetramethylbenzidine
  • Microporous membranes have proven to be highly successful surfaces for the development of simple, rapid immunoassay delivery systems. Membranes made of nitrocellulose can be produced with a wide range of pore sizes and treated with surfactant to optimize their performance. These membranes help create sensitive, stable, and reproducible assays.
  • the key membrane characteristics include polymer type, porosity, surfactant content, and strength. And the choice of a membrane depends largely on three properties: protein binding capacity, porosity, and strength.
  • Polymers tested for their ability to bind proteins include cellulose (nitrocellulose, cellulose acetate, regenerated cellulose), nylon, and polyvi ⁇ ylidene fluoride (PVDF).
  • the binding capacities of cellulose acetate and regenerated cellulose are insufficient for most immunoassays, but a range of higher capacities are available with nitrocellulose, nylon and PVDF membranes.
  • Nitrocellulose is the membrane of choice, even though the protein binding capacity is lower than that of nylon, and it can be manufactured in a wide range of porosities, from 0.05 to about 12 ⁇ m, providing a range of binding capacities and wicking or capillary-rise rates to aid in the flow of reactants through the assay.
  • the modified HIV antigens of the present invention can be used membrane assays.
  • colloidal gold particles sold by BBI Reagents
  • the adsorbed receptor will bind to the biotin ligand group attached to the HIV antigens leaving the HIV epitopes exposed.
  • the receptor molecules can also be attached to the membrane either by adsorption or by covalent binding, by techniques know to those of skill in the art. After adsorption the receptors will bind to the ligand attached to the HIV antigens leaving the HIV epitopes exposed.
  • System 100 comprises a membrane strip 125, a sample pad 110 for blood application, a conjugate pad 120 which has multiple HIV epitopes coated on colloid gold particles, a test region 130 with fixed multiple HIV epitopes, a control region 140 with an animal antibody to HIV antigen, and a reservoir pad 150.
  • the sample pad 110 may be obtained from commercial sources. Suitable sample pads include those manufactured by Millipore and sold as part no. AP-10.
  • the reservoir pad 150 may be formed of the same material as the sample pad.
  • the conjugate pad may be formed as follows: The conjugate pad (Millipore part no. QR01-000-005) is placed in a solution of colloidal gold particles bearing the p24, gp36 and gp41 HIV antigens (available from BBI International). The colloidal gold particles are preferably in a concentration of about 300 OD per 400 cm 2 . The conjugate pad is submerged in the solution and placed on a rocker for about 30 minutes. The conjugate pad is then removed, placed on a piece of aluminum foil and put into a freezer maintained at -60°C for 4 to 16 hours. The pad is removed and cut into strips, which are then i ⁇ ophilized. The strips may be stored at 15-30°C until needed.
  • the membrane strip may be prepared as follows: a 6 mg/mL solution of Mouse anti-p24 antibody in 10mM phosphate buffer is dispensed at 1.8 microiiters/cm as a stripe along the nitrocellulose membrane (Millipore part no.
  • HIV antigens p24, gp36 and gp41, at a concentration of 2 mg/mL for each antigen are dispenses at 4 microliters/cm as a test line.
  • the membrane may be stored dessicated at 15-30°C until needed.
  • the gold particles in conjugate pad 120 are prepared by adsorbing avidin onto the particles. Suitable gold particles may be obtained from BBI Reagents or E.Y. Laboratories, Inc., and the avidin may be adsorbed to these particles. Biotin labeled HIV epitopes are then added and become bound to the particles through the bioti ⁇ -avidin reaction leaving the HIV epitopes exposed.
  • sample pad 100 To perform an HIV assay with system 100, a sample of blood plasma to be tested is placed on sample pad
  • An elutio ⁇ buffer is used to facilitate migration of the sample.
  • a suitable elutio ⁇ buffer may comprise 8.5g sodium chloride, 0.16g monobasic sodium phosphate, 1.24g dibasic sodium phosphate, 5 mL Tween 20, dissolved in water to a total volume of 1.0 L and adjusted to a pH of 7.2-7.6.
  • Plasma migrates through conjugate pad 120 and membrane 125 by capillary action. Any anti-HIV antibodies in the sample will the antibodies react with the HIV epitopes on the gold particles when the antibodies move through the conjugate pad 120.
  • Anti-HIV antibodies will bind to the HIV antigens linked to the gold particles, and the gold particle also moves along the membrane. Many of the attached anti-HIV antibodies have one arm of the antibody molecule free for additional binding.
  • test region 130 When the antibody-antigen-particle complex reaches test region 130, this free antibody binding site reacts with the fixed HIV antigens attached to the membrane and the whole complex becomes attached. The accumulation of large members of gold particles in test region 130 results in a red colored band. Excess particles will continue on and will be bound by anti-HIV antibody prepared in animals and will appear as a red band in the control region 140.
  • the test result is interpreted as follows: The presence of two visible bands, one in the test region and one in the control region, denotes a "positive" result for HIV infection. The presence of only a single line in the control region denotes a "negative" result for HIV infection. Test results are preferably obtained in 5 minutes or less, more preferably 3 minutes or less, and optimally 1-2 minutes.

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Abstract

L'invention concerne des antigènes du VIH et des récepteurs correspondants qui peuvent être incorporés dans des dosages d'anticorps anti-VIH existants, ce qui augmente de manière avantageuse la sensibilité et l'exactitude des dosages. Dans un mode de réalisation, des antigènes VIH sont biotinylés, et le récepteur correspondant est une molécule d'avidine.
PCT/US2000/019017 1999-07-12 2000-07-12 Dosages ameliores de detection d'infection au virus de l'immunodeficience humaine WO2001003721A1 (fr)

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AU62111/00A AU6211100A (en) 1999-07-12 2000-07-12 Improved assays for detecting human immunodeficiency virus infection

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US35171799A 1999-07-12 1999-07-12
US09/351,717 1999-07-12

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Cited By (6)

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Publication number Priority date Publication date Assignee Title
EP1898216A2 (fr) 2006-08-28 2008-03-12 Sysmex Corporation Réactif de diagnostic et kit de réactif de diagnostic d'infection par VIH, procédé pour la fabrication du réactif de diagnostic d'infection par VIH et procédé pour détecter l'anticorps anti-VIH
CN103389373A (zh) * 2012-05-08 2013-11-13 北京勤邦生物技术有限公司 一种检测磺胺类药物的试纸及其应用
JP2018510354A (ja) * 2015-03-30 2018-04-12 ハイコア バイオメディカル エルエルシー 自己免疫および感染性疾患の診断アッセイを実施するための自動免疫分析システム
CN109324191A (zh) * 2008-09-24 2019-02-12 菲鹏生物股份有限公司 间接标记纳米颗粒的抗体检测捕获法及其试剂盒
US10732110B2 (en) 2013-03-15 2020-08-04 Hycor Biomedical, Llc Automated immunoanalyzer system for performing diagnostic assays for autoimmune and infectious diseases
WO2021072593A1 (fr) * 2019-10-14 2021-04-22 深圳迈瑞生物医疗电子股份有限公司 Kit de réactifs et procédé d'utilisation dans la détection d'anticorps anti-vih

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