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WO1992007961A1 - Detection specifique d'anticorps contre les virus du syndrome immunodeficitaire acquis - Google Patents

Detection specifique d'anticorps contre les virus du syndrome immunodeficitaire acquis Download PDF

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Publication number
WO1992007961A1
WO1992007961A1 PCT/US1991/007802 US9107802W WO9207961A1 WO 1992007961 A1 WO1992007961 A1 WO 1992007961A1 US 9107802 W US9107802 W US 9107802W WO 9207961 A1 WO9207961 A1 WO 9207961A1
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htlv
leu
pro
thr
ser
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PCT/US1991/007802
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Myron E. Essex
Yi-Ming A. Chen
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President And Fellows Of Harvard College
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Priority to JP4500779A priority Critical patent/JPH06503170A/ja
Publication of WO1992007961A1 publication Critical patent/WO1992007961A1/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
    • C12N2740/14011—Deltaretrovirus, e.g. bovine leukeamia virus
    • C12N2740/14022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • This invention generally relates to peptides, methods, and kits for detecting antibodies to Human T- Cell Leukemia Viruses (HTLV's) or to HTLV-infected cells in a biological sample.
  • HTLV Human T- Cell Leukemia Viruses
  • HTLV-II Human T- Cell Leukemia Viruses
  • HTLV-III Human T- Cell Leukemia Viruses
  • HTLV-I Human T-cell leukemia virus type I
  • ATLL human adult T-cell leukemia and lymphoma
  • HTLV-I-associated neurological disorder termed HTLV-I-associated
  • HAM/TSP myelopathy/tropical spastic paraparesis
  • HTLV-I infection is endemic to Southeastern Japan and the Caribbean islands (P.H. Levine et al., Int . J. Cancer
  • HTLV-II Human T-cell leukemia virus type II
  • HCL hairy cell leukemia
  • glycoprotein termed gp67.
  • glycoprotein (gp61).
  • HTLV-I or HTLV-II there are various tests to determine the presence of antibodies to HTLV-I or HTLV-II in a biological specimen (Particle Agglutination Assay, Fujirebio Inc., Tokyo, Japan; EIA, Abbott Laboratories, North Chicago, IL; HTLV-I EIA, Dupont Company, Wilmington, DE).
  • an ELISA test is used for blood bank screening.
  • disrupted HTLV-I virions or recombinant HTLV-I virion proteins are used as the source of antigen for the ELISA test.
  • This assay detects antibodies to HTLV-I and, at least, in some cases, also detects antibodies to HTLV-II. H. Lee et al.
  • HTLV-I-encoded markers may fail to detect a non-trivial percentage of HTLV-II-positive samples.
  • the invention features a method for detecting in a biological sample an antibody to a virion of a human T-cell leukemia virus (HTLV) or to a cell infected with the HTLV.
  • the method involves providing an antigen encoded by HTLV-type II (HTLV-II), contacting a biological sample with the antigen, and detecting
  • the invention features a test kit for detecting in a biological sample an antibody to a virion of a human T-cell leukemia virus (HTLV) or to a cell infected with the HTLV.
  • HTLV human T-cell leukemia virus
  • HTLV-II HTLV type II
  • a second container containing a means.for detecting the formation of an immunocomplex between the antibody and the antigen.
  • the antigen is preferably all, or an antigenic segment or analog, of the HTLV-II envelope protein
  • HTLV-II envelope protein preferably the carboxy-terminal half of the HTLV-II envelope protein; more preferably, RP-IIB or a segment or analog thereof which is reactive with RP-IIB antibodies; and, most preferably, RP-B2 or a segment or analog thereof which is reactive with RP-B2 antibodies.
  • RP-IIB or a segment or analog thereof which is reactive with RP-IIB antibodies preferably the carboxy-terminal half of the HTLV-II envelope protein
  • RP-IIB or a segment or analog thereof which is reactive with RP-IB antibodies preferably, RP-B2 or a segment or analog thereof which is reactive with RP-B2 antibodies.
  • the method further involves screening for both HTLV-I and HTLV-II.
  • the HTLV-II-encoded marker In addition to the use of the HTLV-II-encoded marker
  • such a screen involves contacting the biological sample with at least one antigen encoded by HTLV-type I (HTLV-I), preferably an envelope segment or analog which is reactive with HTLV-I sera and not with HTLV-II sera, more preferably, RB-B1 or a segment or analog thereof which is reactive with RP-B1 antibodies.
  • HTLV-I HTLV-type I
  • the antigen encoded by HTLV-II and the antigen encoded by HTLV-I may be combined and contacted with the biological sample simultaneously; or they may be maintained in separate containers and contacted with the sample
  • Detection of antigen-antibody complexes preferably is accomplished using a Western blot or an ELISA format.
  • the invention features a method for discriminating between HTLV-I and HTLV-II infection.
  • the method involves: providing HTLV-I recombinant protein RP-B1 or an antigenic segment or analog thereof,
  • the invention features a test kit for detecting in a biological sample an antibody to human T-cell leukemia virus type I (HTLV-I).
  • the kit is compartmentalized to receive in close confinement therein one or more containers which include: a first container containing HTLV-I recombinant protein RP-Bl or an
  • Detection of the immunocomplexes is preferably
  • the invention also features the RP-IIB peptide or a segment or analog thereof, preferably RP-B2, which is reactive with RP-IIB antibodies and, preferably, which is not reactive with antibodies specific for RP-B1; and purified nucleic acid encoding the RB-IIB peptide or a segment or analog thereof which is reactive with RP-IIB antibodies.
  • nucleic acid is included in plasmid, pIIB, or, more preferably, pB2.
  • antigen is meant a substance, in this case, a peptide which interacts in a demonstrable selective way with an antibody (e.g., an antibody which is included in a biological sample) which results from challenge by all or part of the virus at issue.
  • an antibody e.g., an antibody which is included in a biological sample
  • a “peptide” is any chain of amino acids, including
  • antigenic segment or “segment of an
  • antigen is meant any portion of an antigenic peptide which interacts in a demonstrable selective way with an antibody directed against the organism bearing the antigen.
  • analog is meant a peptide differing from the antigen or antigenic segment by one or more
  • a suitable analog as used herein, is one which interacts in a demonstrable selective way with an antibody directed against the organism bearing the antigen.
  • Such analogs may be produced by any method of recombinant DNA
  • HTLV-I-encoded antigen or "HTLV-II- encoded antigen”
  • HTLV-II-encoded antigen we include not only peptides whose amino acid sequence is encoded by a naturally occurring HTLV, but also analogs of such peptides as described above.
  • purified is meant substantially isolated from other cellular components, e.g., proteins, lipids, and other nucleic acids, with which the substance
  • FIG. 1 is the nucleic acid and corresponding amino acid sequence of the HTLV-II provirus (Shimotohno et al., Proc. Natl . Acad . Sci . USA 82:3101, 1985; SEQ ID NO.:1).
  • FIG. 2 is a hydrophobicity plot of the HTLV-II- encoded envelope protein sequence and a comparison of the amino acid sequences of the HTLV-II and HTLV-I envelope proteins.
  • FIG. 3 is the nucleic acid and corresponding amino acid sequence of HTLV-I including the sequence encoding RP-B1 (Seiki et al., Proc. Natl . Acad . Sci . USA 80:3618. 1983; SEQ ID NO. :3).
  • FIG. 4 is a diagram depicting construction of expression plasmids, pB, pB1, and pD.
  • FIG. 5 is a diagram depicting construction of expression plasmids, pA and pC.
  • FIG. 6 is a diagram depicting construction of expression plasmid, pIIB.
  • HTLV- II-encoded serologic markers other than those disclosed below may be used to detect HTLV-II infection according to the invention. These serologic markers would preferably be envelope protein peptide segments or analogs, but may be any HTLV-II-encoded peptide which is reactive with antibodies to HTLV-II. To produce such peptide segments or analogs, HTLV-II-derived nucleic acid encoding putative antigenic peptides may be cloned into an expression vector (e.g., an expression vector
  • peptides may be synthesized by other techniques, including, without limitation, any technique of recombinant DNA technology or chemical, e.g., peptide, synthesis.
  • MO mammalian cell line
  • HTLV-II the mammalian cell line
  • Viral DNA may be isolated from such a cell line by standard techniques
  • HTLV-II DNA may be obtained from available full-length or partial HTLV-II proviral clones (e.g., ⁇ H6.0; Chen et al.. Nature 305:502, 1983; Shimotohno et al., Proc. Natl . Acad. Sci . USA 82:3101, 1985).
  • the full-length HTLV-II sequence is shown in Fig. 1 (SEQ ID NO.:1)
  • ydrophilic regions are more likely to be antigenic, i.e., reactive with HTLV-II-positive sera, because these regions are exposed on the surface of the virion or virion-infected cell.
  • a hydrophobicity plot of the HTLV- II envelope protein sequences is shown in Fig. 2.
  • candidate peptides including peptide segments or analogs
  • Fig. 2 also shows the regions of conserved amino acid residues along the HTLV-I and HTLV-II amino acid
  • Candidate peptides may be tested, by methods described in greater detail below, using, e.g., PCR- confirmed HTLV-II-positive antiserum (as described below).
  • Antigenic peptides are those which react with HTLV-II-positive serum (as determined using the methods described below).
  • Peptides specific for HTLV-II are those which react exclusively or selectively (e.g., a "two plus antibody reactivity", as defined below) with HTLV-II-positive serum.
  • a particularly preferred HTLV-II-encoded antigen is the recombinant protein, RP-IIB (described below; SEQ. ID NO.: 2) or segments or analogs thereof.
  • RP-IIB recombinant protein
  • Candidate segments or analogs may be produced, e.g., by any combination thereof.
  • RP-IIB is included in the peptide RP-B2 (described below).
  • HTLV-I-encoded antigens can be used to detect HTLV infection.
  • the HTLV-I-encoded antigen can be used in addition to the above-described HTLV-II- encoded antigen to reduce the incidence of false negative readings.
  • RP-B1 is desirable to use RP-B1, as described below, or a segment or analog thereof.
  • Candidate segments or analogs of RP-B1 would be generated and screened for antigenicity as described above for RP-IIB, and for lack of reactivity with HTLV-II.
  • Sources of HTLV-I include the mammalian cell lines, termed MJ and C5-MJ, which are permanently and persistently infected with HTLV-I; these cell lines are available from the American Type Culture Collection (ATCC Accession Nos. CRL-8294 and CRL-8293, respectively).
  • Viral DNA may be isolated from such cell lines by standard techniques (see, e.g., Tsujimoto et al., Mol . Biol . Med. 5:29, 1988; Seiki et al., Proc .
  • HTLV-I DNA may be obtained from available full-length or partial HTLV-I proviral clones (e.g., ⁇ ATM-1 or ⁇ ATK-1; Seiki et al., Proc. Natl . Acad. Sci . USA 80.3618, 1983).
  • the HTLV-1 sequence encoding RP-B1 is shown in Fig. 3 (SEQ ID NO.: 3) Experimental Information
  • the preferred HTLV serologic markers according to the invention are identified from a panel of recombinant proteins containing different regions of the HTLV-I or HTLV-II env proteins whose immunological reactivity is assayed as described below.
  • fragments of HTLV-I proviral DNA sequences were cloned into appropriate expression vectors, using as a
  • the intact env gene was carried on the plasmid, psphl-envl.
  • This plasmid was constructed as follows.
  • the plasmid pMT2 (Clark et al., Nature 305:60, 1983, hereby incorporated by reference) was digested with SphI and a 4.2-kilobase (kb) fragment containing both the HTLV-1 env gene and the x region was isolated and subcloned into expression vector, p806 (a derivative of pUC18 containing the tac promoter and v- ras H gene on an EcoRI-NcoI fragment which was originally derived from plasmid pXVR; Feig et al., Proc . Natl . Acad. Sci USA 83: 4607, 1987, hereby incorporated by reference).
  • the inserted env gene was determined by sequence analysis (by the procedure of Toneguzzo et al., Biotechniques
  • the distal two-thirds of the HTLV-I env gene was expressed from plasmid, pS3.
  • psphl- envl was digested with Sall, and a SalI-ended fragment containing the first 5694 nucleotides of the env gene was deleted.
  • the resultant plasmid (containing nucleotides 5694 to 6665 of the env gene) was treated with DNA polymerase Klenow fragment and religated by blunt-end ligation (Fig. 4).
  • the transmembrane env protein (encoded by nucleotides 6140 to 6665), was expressed from plasmid pD (Fig. 4).
  • psphl-envl DNA was digested with Kpnl , and a kpnl-ended fragment containing the first 6140 nucleotides of the env gene was deleted.
  • the plasmid DNA (containing nucleotides 6140 to 6665 of the env gene) was treated with DNA polymerase Klenow fragment and religated by blunt end ligation.
  • plasmids Two internal regions of the HTLV-I env gene were expressed from plasmids, pB and pBl (Fig. 4). These plasmids were generated from plasmid pS3, isolated from E. coli strain JM110 and digested with either Clal or Xhol . Ends were blunted with DNA polymerase Klenow fragment and re-ligated, by blunt-end ligation, in the presence of Nhel nonsense codon linker DNA,
  • the resultant plasmids, pB and pBl contained HTLV-1 env gene nucleotide sequences (5694 to 5887) and (5694 to 5799; SEQ ID NO.: 3), respectively (Fig. 4).
  • Expression vector pJL6 (Lautenberger et al., Gene Anal . Technol . 42: 49 , 1984, hereby incorporated by reference) was used to express the N-terminal half of the HTLV-I envelope glycoprotein, gp46.
  • pJL6 contains a bacteriophage ⁇ ⁇ L promoter and the N-terminal fragment of the ⁇ cll gene, including the ribosome-binding site and an ATG start codon.
  • bp 422-base- pair
  • Plasmid pC (Fig. 5), encoding the C-terminal region of gp46, was
  • the recombinant protein, RP-IIB was expressed from plasmid, pIIB.
  • This plasmid was constructed by isolating, from plasmid pMO1A (Gelmann et al., Proc.
  • pMO1A may be digested, as described above, with Rsal and a 420 base pair fragment isolated for insertion into the desired vector.
  • Recombinant proteins containing specific regions of HTLV-1 gp61 were produced by either vector p806 (pB, pB1, and pD) or by vector JL6 (pA and pC) in two different bacterial culture systems.
  • E. coli X-90 (Pallas et al., J. Virol . 40:1075, 1986, hereby
  • the RPs were partially purified as described in Matsuda et al. (Proc. Natl . Acad . Sci . USA 85:6968, 1988) and were identified by Coomassie blue staining (as described in Reisner et al., Anal . Biochem . 64: 509, 1975, hereby incorporated by reference) of cellular proteins which had been separated by electrophoresis on a sodium dodecyl sulfate-polyacrylamide (SDS-PA) gel (Laemmli, Nature 227:680, 1979). Their molecular weight (in kDa) was determined by comparison with protein standards (of known molecular weight); the apparent molecular weight of each RP is given in Table 1.
  • N N terminus
  • M middle region
  • C C terminus of gp46, the exterior domain of gp61
  • gp21 the transmembrane domain of gp61.
  • the pJL6 bacterial system generally produced the RPs in greater quantity than did the p806 system, however, the reason for this difference is unclear.
  • RP-D Western blot
  • RP-IIB was produced by transformation of pIIB into E. coli strain DC1148.
  • An 18 kD protein was expressed by pIIB and was shown to be reactive to an HTLV-II-positive serum by WB assay 20 minutes after the temperature shift. Its production reached a plateau 40 minutes after
  • the bacterial pellet was subjected to a series of partial purification steps which included successive high salt and detergent extractions combined with sonication as described in Matsuda et al. (Proc. Natl . Acad . Sci . USA 85:6968,
  • RP-IIB was subsequently detected in an 8M urea fraction by Coomassie blue staining of an SDS- polyacrylamide gel (Laemmli, Nature 227:680. 1979, hereby incorporated by reference).
  • Serum samples from two groups of Japanese HTLV-1- positive subjects, healthy carriers, and ATL patients were used to study the prevalence of antibodies to each of the specific RPs.
  • Initial screening for HTLV-1 seropositivity was standardized by using both the particle agglutination test (Ikeda et al., Gann . 75:845, 1984, hereby incorporated by reference) and the
  • antibody reactivity to the RPs can be summarized as follows: 22.2% (10 of 45) of samples from the carriers and 16.9% (13 of 77) of samples from the ATL patients were reactive to RP-A; 80% (36 of 45) of samples from the carriers and 68.8% (53 of 77) of the samples from the ATL patients were reactive to RP- B1; and 91.1% (41 of 45) of samples from the carriers and 93.8% (76 of 81) of samples from the ATL patients were reactive to RP-C.
  • RP-A (P ⁇ 0.005) than that detected by the N-terminal half of gp46 (i.e., RP-A).
  • RP-A, RP-B, and RP-C which together span the entire length of gp46 except the first five amino acids at the N-terminus and the last four amino acids at the C-terminus, detected 99.2% (125 of 126) of the HTLV-I-positive subjects. This same combination of RPs detected every ATL patient in this study.
  • RP-D which contains the transmembrane envelope protein gp21 minus the first amino acid at the N- terminus, had only a 73.7% (84 of 114) reactivity rate.
  • Recombinant Protein B2 contains an HTLV-II-specific
  • pB2 To produce expression plasmid, pB2, a DNA fragment including nucleic acid residues 5675 to 5801 was ligated, by blunt end ligation, into Smal-/ Sphl-digested p806; both the RP-B2-encoding fragment and the p806 vector were treated with Klenow fragment to produce blunt ends prior to ligation.
  • the 0.16 kb RP-B2-encoding fragment was produced by standard methods of PCR amplification, using plasmid pIIB as a template and the primers:
  • Plasmid pB2 encodes a fusion protein, termed RP-B2, containing 48 amino acids from the HTLV-II envelope protein (i.e., amino acid residues 166 to 213 of Fig. 1; SEQ ID NO.: 1).
  • RP-B2 showed specificity for HTLV-II-positive sera in WB assays. None of the 12 HTLV-I-positive sera, shown above to cross-react with RP-IIB, had detectable antibody reactivity to RP-B2; all 20 HTLV-II-positive sera
  • Recombinant Protein Bl contains an HTLV-I-specific
  • HTLV-II-positive serum samples from intravenous drug abusers from New Orleans; Serologicals, Inc., Marietta, GA
  • PCR confirmation was carried out by analyzing isolated lymphocyte DNA according to the 32 [P]-oligonucleotide end- labelling PCR technique described previously (Lee et al., Science 244:471.1989, hereby incorporated by reference)
  • Five oligonucleotide primer pairs corresponding to tax, protease and LTR sequences were utilized. The tax oligonucleotide primers corresponded to conserved
  • RPs were analyzed for antibody reactivity by Western blotting (WB) as described above. Partially purified RPs solubilized either in 3M or 8M urea were subjected to electrophoresis on a 15% SDS- polyacrylamide gel (Laemmli, Nature 227:680. 1970 and passively transferred to nitrocellulose membranes
  • Antibody reactivity with each of the RPs were determined to be as follows: 10% (2/20) for RP-A; 5%
  • RP-IIB which contained amino acid residues 96-235 from the HTLV-II exterior envelope glycoprotein (i.e., amino acid residues 21-308) was reactive to all 20 HTLV-II serum samples.
  • RP- Bl and RP-IIB proteins were used in a Western blot assay to study the antibody reactivity of 115 additional plasma samples. Those samples included: 9 PCR-confirmed HTLV- I and 45 PCR-confirmed HTLV-II plasma samples from intravenous drug abusers in New Jersey, 13 HTLV-I-Enzyme immunoassay negative samples, l HTLV-I-WB indeterminate case, and 1 HTLV-I seropositive but PCR-negative case. Another panel of 46 PCR-confirmed HTLV-I plasma samples were selected from a nationwide survey of food service employees in 1987-1988 in Jamaica. The samples were randomly mixed, blind-coded by a collaborator from
  • Recombinant protein IIB contains HTLV-II-specific
  • HTLV-I carriers were confirmed by PCR according to the technique of Lee et al. (Proc. Natl . Acad. Sci . USA 81 : 1519 , 1984) described above. Among 27 HTLV-II carriers confirmed by PCR, two carriers were seronegative in Abbott and Du Pont HTLV-I EIAs and had indeterminate antibody profile for HTLV-I infection
  • the intensity of reactivity to RP-IIB was classified into +, ++ and +++ according to the O.D.
  • the optic density (O.D.) of reactive bands in the WB results was recorded by densitometer scanning (Model 620, BioRad Laboratories, Richmond, CA) and scored one plus to three plus according to a panel of standard reactivities which were tested side by side with the test samples in WB assays.
  • Anti-RP-IIB antibody reactivities of a standard PCR-confirmed HTLV-II serum were tested at 1:200, 1:2,000 and 1:10,000 dilutions.
  • the score was one plus; for serum reactivity equal to the standard serum at 1:2,000 or between those of the standard serum at 1:10,000 and 1:2,000, the score was two plus; and the score was three plus when a sample had antibody reactivity greater than that of the standard serum at 1:2000 dilution. This data is also summarized in Table 4.
  • HTLV-II serum samples had two or three plus seroreactivity to RP-IIB while, among those HTLV-I positive samples cross-reacting to RP-IIB, only 1 of 14 (7%) had two plus reactivity. This was also reflected by the significant difference in the means of O.D. from densitometer scanning for the above two groups, i.e., 1.19 ⁇ 0.11 for HTLV-II samples and 1.04 ⁇ 0.05 for HTLV-I samples (t-test, p ⁇ 0.05); Statistical Methods in the Biological and Health Sciences, ed., Milton and
  • Antibody reactivity may be recorded using any of a number of immunoassays well known to those skilled in the art.
  • the antigenic peptide can be radio-labelled by conventional methods for use in radioimmunoassay, with fluorescein for fluorescent immunoassay, with enzyme for enzyme immunoassay, or with biotin for biotin-avidin linked assays, it can be employed, labelled or
  • the antigenic peptide can also be immobilized on some solid phase, such as an insoluble resin (e.g., a nitrocellulose filter) or a microtiter well, and detection of the HTLV antibody carried out by measuring binding of the antibody to the solid phase.
  • Solid phases may also include latex particles, which, when coated with the antigenic peptide and subjected to reactive antibody, will agglutinate.
  • antigenic peptide may be attached in various examples.
  • solid phases to which the antigenic peptide may be attached include, without limitation, test tubes, vials, titration wells, and the like.
  • Antibody may be detected by double antibody techniques or Protein- A dependent techniques.
  • test kits are incorporated into test kits. Such kits are
  • the first compartment includes one or more of the antigenic peptides of the invention, in detectably labelled form (e.g., any of the forms above) or immobilized on a solid phase (e.g., those listed above).
  • the second container may include elements necessary for detection of the label on the antigen
  • a second antibody e.g., monoclonal or polyclonal anti IgG directed to the antibody in the biological sample which is specific for the HTLV-encoded antigen.
  • a second antibody e.g., monoclonal or polyclonal anti IgG
  • CTAGACTCTG CCTTAAACTT CACTTCCGCG TTCTTGTCTC GTTCTTTCCT CTTCGCCGTC 600
  • CAA CTC AAA CCC CCT CAG GAG GAA GGG GAA CCC CTC CTG TTG GAT CTC 2054 Gln Leu Lys Pro Pro Gln Glu Glu Gly Glu Pro Leu Leu Leu Asp Leu
  • AAA AAC ACA ACA AAC CCC AGG CCA AAT ACG CTT CTT AGG ACA GGT CAT 3206 Lys Thr Gln Gln Thr Pro Gly Gln Ile Arg Phe Lue Gly Gln Val Ile
  • GGC CTT GCC ACC CCT ACT GCA GGG CAA GAC CAT CTA CCT CCA CCA TGT 4214 Ala Leu Pro Pro Leu Leu Gln Gly Lys Thr Ile Tyr Leu His His Val
  • CAC AGA CTC CCT TAT CTT AGC TCC CCT TGT TCC CCT GAC GCC CCA AGG 4310 Thr Asp Ser Leu Ile Leu Ala Pro Leu Val Pro Leu Thr Pro Gln Gly
  • GCA GCC CAA AAT AGA CGA GGA TTA GAC CTC CTA TTC TGG GAA CAA GGG 6337 Ala Ala Gln Asn Arg Arg Gly Leu Asp Leu Leu Phe Trp Glu Gln Gly
  • GTC ATC ACC GGC TGG GGA CTA AAC TGG GAT CTT GGA CTG TCC CAA TGG 6481 Val Ile Thr Gly Trp Gly Leu Asn Trp Asp Leu Gly Leu Ser Gln Trp
  • GCA GAA GAA GCC CTC CAG ACA GGC ATA ACC ATT CTC GCT CTA CTC CTC 6529 Ala Arg Glu Ala Leu Gln Thr Gly Ile Thr Ile Leu Ala Leu Leu Leu Leu
  • TCCACAGTCC TCTATACCAG ATGAGTCGCC CCCGATGTCC AGCCCTAACT CGATTCTGAA 6757
  • ATC CCT GTC TCT ATT TTA TTT AAT AAA GAA GAG GCG GAT GAC AAT GGC 8199 Ile Pro Val Ser Ile Leu Phe Asn Lys Glu Glu Ala Asp Asp Asn Gly
  • CAA GAC ATC TCG ATA CTC CCA CTC ATC CCC CTG CGG CAG CAA CAG CAA 9048 Gln Asp Ile Ser Ile Leu Pro Leu Ile Pro Leu Arg Gln Gln Gln Gln Gln

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Abstract

Procédé et trousse de test pour la détection dans un échantillon biologique d'un anticorps contre un virion d'un virus du syndrome immunodéficitaire acquis (HTLV) ou contre une cellule infectée par un tel virion. Le procédé consiste généralement à mettre en contact l'échantillon biologique avec un antigène codé par HTLV-II et à détecter par essai immunologique la formation du complexe antigène-anticorps. On décrit également un procédé apparenté et une trousse de test pour la détection d'un virion de HTLV-I ou d'une cellule infectée par HTLV-I; ce procédé consiste généralement à mettre en contact l'échantillon biologique avec RP-B1, l'antigène spécifique de HTLV-I, et à détecter par essai immunologique la formation du complexe antigène-anticorps. On décrit également les peptides RP-IIB et RP-B2 codés par HTLV-II et les acides nucléiques purifiés codant ces peptides.
PCT/US1991/007802 1990-10-26 1991-10-22 Detection specifique d'anticorps contre les virus du syndrome immunodeficitaire acquis WO1992007961A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP4500779A JPH06503170A (ja) 1990-10-26 1991-10-22 ヒトｔ細胞白血病ウイルスに対する抗体の特異的検出

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US60284690A 1990-10-26 1990-10-26
US602,846 1990-10-26

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WO1992007961A1 true WO1992007961A1 (fr) 1992-05-14

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EP (1) EP0555405A4 (fr)
JP (1) JPH06503170A (fr)
AU (1) AU8944691A (fr)
CA (1) CA2094832A1 (fr)
WO (1) WO1992007961A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990010231A1 (fr) * 1989-03-02 1990-09-07 Replico Medical Ab Discrimination entre des anticorps contre htlv-i, htlv-ii ou des retrovirus apparentes, des nouveaux peptides, detection d'anticorps et kits d'immunoanalyse
US5017687A (en) * 1988-03-10 1991-05-21 Virovahl, S.A. Peptides for the detection of HTLV-1 infection

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0196319B1 (fr) * 1984-09-19 1992-01-02 The Regents Of The University Of California Polypeptides retroviraux associes a la transformation de cellules humaines
WO1989006543A1 (fr) * 1988-01-12 1989-07-27 Genelabs Incorporated Antigene peptidique de l'htlv-i et procede d'analyse utilisant un tel antigene
FI910245L (fi) * 1990-01-24 1991-07-25 United Biomedical Inc Syntetiska peptidkompositioner med immunoreaktivitet mot htlv-antikroppar.
JPH04164097A (ja) * 1990-10-24 1992-06-09 Kuraray Co Ltd ペプチドおよびその用途

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5017687A (en) * 1988-03-10 1991-05-21 Virovahl, S.A. Peptides for the detection of HTLV-1 infection
WO1990010231A1 (fr) * 1989-03-02 1990-09-07 Replico Medical Ab Discrimination entre des anticorps contre htlv-i, htlv-ii ou des retrovirus apparentes, des nouveaux peptides, detection d'anticorps et kits d'immunoanalyse

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0555405A4 *

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JPH06503170A (ja) 1994-04-07
EP0555405A4 (en) 1993-08-25
EP0555405A1 (fr) 1993-08-18
AU8944691A (en) 1992-05-26
CA2094832A1 (fr) 1992-04-27

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