Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 May;27(5):977–982. doi: 10.1128/jcm.27.5.977-982.1989

Reliable confirmation of antibodies to human immunodeficiency virus type 1 (HIV-1) with an enzyme-linked immunoassay using recombinant antigens derived from the HIV-1 gag, pol, and env genes.

V L Ng 1, C S Chiang 1, C Debouck 1, M S McGrath 1, T H Grove 1, J Mills 1
PMCID: PMC267466  PMID: 2787334

Abstract

An enzyme-linked immunoassay (ELISA) using six recombinant proteins corresponding to large segments of the human immunodeficiency virus type 1 (HIV-1) gag, pol, and env gene products (HIVAGEN; SmithKline Bio-Science Laboratories, Van Nuys, Calif.) was developed to confirm the presence of antibodies to HIV-1 in sera reactive in the whole-cell-derived virion screening ELISAs. Serum samples for testing were obtained from healthy seronegative blood donors and from the different categories of HIV-infected individuals (asymptomatic, acquired immunodeficiency syndrome [AIDS]-related complex, and AIDS). A positive reaction was defined as reactivity against an env and at least one other (either gag or pol) HIV-1 gene product; negative was defined as no reaction with any antigen; and indeterminate was defined as reactivity with gag or pol (or both) or with env alone. None of the 1,180 serum samples from healthy seronegative blood donors gave a positive result, and only 49 of these samples (4%) gave indeterminate results. The recombinant HIV-1 antigen ELISA panel identified seropositive individuals with a high degree of accuracy, as a positive reaction was seen with 99.3% of asymptomatic healthy seropositive individuals, 98.1% of patients with AIDS-related complex, and 90.4% of patients with AIDS. None of the 725 HIV-1-seropositive subjects had a negative test result. Reactivity with the Kp41 antigen, corresponding to an amino-terminal portion of the gp41 envelope glycoprotein, by itself demonstrated 100% sensitivity and specificity in distinguishing seronegative from seropositive sera. A subset of seronegative and seropositive samples were tested both with the recombinant HIV-1 antigen ELISA panel and by Western blot (Du Pont Co.). The recombinant HIV-1 antigen ELISA panel accurately identified more seropositive and seronegative samples and had fewer indeterminate results than did Western blot (interpreted by Du Pont criteria).

Full text

PDF
977

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allain J. P., Laurian Y., Paul D. A., Verroust F., Leuther M., Gazengel C., Senn D., Larrieu M. J., Bosser C. Long-term evaluation of HIV antigen and antibodies to p24 and gp41 in patients with hemophilia. Potential clinical importance. N Engl J Med. 1987 Oct 29;317(18):1114–1121. doi: 10.1056/NEJM198710293171804. [DOI] [PubMed] [Google Scholar]
  2. Barré-Sinoussi F., Chermann J. C., Rey F., Nugeyre M. T., Chamaret S., Gruest J., Dauguet C., Axler-Blin C., Vézinet-Brun F., Rouzioux C. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science. 1983 May 20;220(4599):868–871. doi: 10.1126/science.6189183. [DOI] [PubMed] [Google Scholar]
  3. Biberfeld G., Bredberg-Rådén U., Böttiger B., Putkonen P. O., Blomberg J., Juto P., Wadell G. Blood donor sera with false-positive Western blot reactions to human immunodeficiency virus. Lancet. 1986 Aug 2;2(8501):289–290. doi: 10.1016/s0140-6736(86)92111-2. [DOI] [PubMed] [Google Scholar]
  4. Burke D. S., Brandt B. L., Redfield R. R., Lee T. H., Thorn R. M., Beltz G. A., Hung C. H. Diagnosis of human immunodeficiency virus infection by immunoassay using a molecularly cloned and expressed virus envelope polypeptide. Comparison to Western blot on 2707 consecutive serum samples. Ann Intern Med. 1987 May;106(5):671–676. doi: 10.7326/0003-4819-106-5-671. [DOI] [PubMed] [Google Scholar]
  5. Carlson J. R., Yee J., Hinrichs S. H., Bryant M. L., Gardner M. B., Pedersen N. C. Comparison of indirect immunofluorescence and Western blot for detection of anti-human immunodeficiency virus antibodies. J Clin Microbiol. 1987 Mar;25(3):494–497. doi: 10.1128/jcm.25.3.494-497.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cooper D. A., Gold J., Maclean P., Donovan B., Finlayson R., Barnes T. G., Michelmore H. M., Brooke P., Penny R. Acute AIDS retrovirus infection. Definition of a clinical illness associated with seroconversion. Lancet. 1985 Mar 9;1(8428):537–540. doi: 10.1016/s0140-6736(85)91205-x. [DOI] [PubMed] [Google Scholar]
  7. Couroucé A. M., Muller J. Y., Richard D. False-positive Western blot reactions to human immunodeficiency virus in blood donors. Lancet. 1986 Oct 18;2(8512):921–922. doi: 10.1016/s0140-6736(86)90443-5. [DOI] [PubMed] [Google Scholar]
  8. Dawson G. J., Heller J. S., Wood C. A., Gutierrez R. A., Webber J. S., Hunt J. C., Hojvat S. A., Senn D., Devare S. G., Decker R. H. Reliable detection of individuals seropositive for the human immunodeficiency virus (HIV) by competitive immunoassays using Escherichia coli-expressed HIV structural proteins. J Infect Dis. 1988 Jan;157(1):149–155. doi: 10.1093/infdis/157.1.149. [DOI] [PubMed] [Google Scholar]
  9. Frösner G. G., Erfle V., Mellert W., Hehlmann R. Diagnostic significance of quantitative determination of HIV antibody specific for envelope and core proteins. Lancet. 1987 Jan 17;1(8525):159–160. doi: 10.1016/s0140-6736(87)91990-8. [DOI] [PubMed] [Google Scholar]
  10. Gallo D., Diggs J. L., Shell G. R., Dailey P. J., Hoffman M. N., Riggs J. L. Comparison of detection of antibody to the acquired immune deficiency syndrome virus by enzyme immunoassay, immunofluorescence, and Western blot methods. J Clin Microbiol. 1986 Jun;23(6):1049–1051. doi: 10.1128/jcm.23.6.1049-1051.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gallo R., Wong-Staal F., Montagnier L., Haseltine W. A., Yoshida M. HIV/HTLV gene nomenclature. Nature. 1988 Jun 9;333(6173):504–504. doi: 10.1038/333504a0. [DOI] [PubMed] [Google Scholar]
  12. Goudsmit J., Lange J. M., Paul D. A., Dawson G. J. Antigenemia and antibody titers to core and envelope antigens in AIDS, AIDS-related complex, and subclinical human immunodeficiency virus infection. J Infect Dis. 1987 Mar;155(3):558–560. doi: 10.1093/infdis/155.3.558. [DOI] [PubMed] [Google Scholar]
  13. Gupta P., Balachandran R., Grovit K., Webster D., Rinaldo C., Jr Detection of human immunodeficiency virus by reverse transcriptase assay, antigen capture assay, and radioimmunoassay. J Clin Microbiol. 1987 Jun;25(6):1122–1125. doi: 10.1128/jcm.25.6.1122-1125.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hofbauer J. M., Schulz T. F., Hengster P., Larcher C., Zangerle R., Kofler H., Fritsch P., Wachter H., Dierich M. P. Comparison of Western blot (immunoblot) based on recombinant-derived p41 with conventional tests for serodiagnosis of human immunodeficiency virus infections. J Clin Microbiol. 1988 Jan;26(1):116–120. doi: 10.1128/jcm.26.1.116-120.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lelie P. N., van der Poel C. L., Reesink H. W. Interpretation of isolated HIV anti-p24 reactivity in Western blot analysis. Lancet. 1987 Mar 14;1(8533):632–632. doi: 10.1016/s0140-6736(87)90274-1. [DOI] [PubMed] [Google Scholar]
  16. Lennette E. T., Karpatkin S., Levy J. A. Indirect immunofluorescence assay for antibodies to human immunodeficiency virus. J Clin Microbiol. 1987 Feb;25(2):199–202. doi: 10.1128/jcm.25.2.199-202.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levy J. A., Hoffman A. D., Kramer S. M., Landis J. A., Shimabukuro J. M., Oshiro L. S. Isolation of lymphocytopathic retroviruses from San Francisco patients with AIDS. Science. 1984 Aug 24;225(4664):840–842. doi: 10.1126/science.6206563. [DOI] [PubMed] [Google Scholar]
  18. Martin P. W., Burger D. R., Caouette S., Goldstein A. S., Peetoom F. Importance of confirmatory tests after strongly positive HTLV-III screening tests. N Engl J Med. 1986 Jun 12;314(24):1577–1577. doi: 10.1056/nejm198606123142412. [DOI] [PubMed] [Google Scholar]
  19. Mayer K. H., Falk L. A., Paul D. A., Dawson G. J., Stoddard A. M., McCusker J., Saltzman S. P., Moon M. W., Ferriani R., Groopman J. E. Correlation of enzyme-linked immunosorbent assays for serum human immunodeficiency virus antigen and antibodies to recombinant viral proteins with subsequent clinical outcomes in a cohort of asymptomatic homosexual men. Am J Med. 1987 Aug;83(2):208–212. doi: 10.1016/0002-9343(87)90686-3. [DOI] [PubMed] [Google Scholar]
  20. Mendenhall C. L., Roselle G. A., Grossman C. J., Rouster S. D., Weesner R. E. False positive tests for HTLV-III antibodies in alcoholic patients with hepatitis. N Engl J Med. 1986 Apr 3;314(14):921–922. doi: 10.1056/NEJM198604033141412. [DOI] [PubMed] [Google Scholar]
  21. Montagnier L., Clavel F., Krust B., Chamaret S., Rey F., Barré-Sinoussi F., Chermann J. C. Identification and antigenicity of the major envelope glycoprotein of lymphadenopathy-associated virus. Virology. 1985 Jul 15;144(1):283–289. doi: 10.1016/0042-6822(85)90326-5. [DOI] [PubMed] [Google Scholar]
  22. Moore J. D., Cone E. J., Alexander S. S., Jr HTLV-III seropositivity in 1971-1972 parenteral drug abusers--a case of false positives for evidence of viral exposure? N Engl J Med. 1986 May 22;314(21):1387–1388. [PubMed] [Google Scholar]
  23. Neale T. J., Dagger J., Fong R., Humble M., Meech P., Hatfield P. J., Morrison R. B. False positive anti-HTLV III serology. N Z Med J. 1985 Oct 23;98(789):914–914. [PubMed] [Google Scholar]
  24. Petricciani J. C. Licensed tests for antibody to human T-lymphotropic virus type III. Sensitivity and specificity. Ann Intern Med. 1985 Nov;103(5):726–729. doi: 10.7326/0003-4819-103-5-726. [DOI] [PubMed] [Google Scholar]
  25. Popovic M., Sarngadharan M. G., Read E., Gallo R. C. Detection, isolation, and continuous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science. 1984 May 4;224(4648):497–500. doi: 10.1126/science.6200935. [DOI] [PubMed] [Google Scholar]
  26. Ranki A., Johansson E., Krohn K. Interpretation of antibodies reacting solely with human retroviral core proteins. N Engl J Med. 1988 Feb 18;318(7):448–449. doi: 10.1056/NEJM198802183180712. [DOI] [PubMed] [Google Scholar]
  27. Saag M. S., Britz J. Asymptomatic blood donor with a false positive HTLV-III Western blot. N Engl J Med. 1986 Jan 9;314(2):118–118. doi: 10.1056/nejm198601093140212. [DOI] [PubMed] [Google Scholar]
  28. Saah A. J., Farzadegan H., Fox R., Nishanian P., Rinaldo C. R., Jr, Phair J. P., Fahey J. L., Lee T. H., Polk B. F. Detection of early antibodies in human immunodeficiency virus infection by enzyme-linked immunosorbent assay, Western blot, and radioimmunoprecipitation. J Clin Microbiol. 1987 Sep;25(9):1605–1610. doi: 10.1128/jcm.25.9.1605-1610.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sayers M. H., Beatty P. G., Hansen J. A. HLA antibodies as a cause of false-positive reactions in screening enzyme immunoassays for antibodies to human T-lymphotropic virus type III. Transfusion. 1986 Jan-Feb;26(1):113–115. doi: 10.1046/j.1537-2995.1986.26186124012.x. [DOI] [PubMed] [Google Scholar]
  30. Schüpbach J., Haller O., Vogt M., Lüthy R., Joller H., Oelz O., Popovic M., Sarngadharan M. G., Gallo R. C. Antibodies to HTLV-III in Swiss patients with AIDS and pre-AIDS and in groups at risk for AIDS. N Engl J Med. 1985 Jan 31;312(5):265–270. doi: 10.1056/NEJM198501313120502. [DOI] [PubMed] [Google Scholar]
  31. Shatzman A. R., Rosenberg M. Expression, identification, and characterization of recombinant gene products in Escherichia coli. Methods Enzymol. 1987;152:661–673. doi: 10.1016/0076-6879(87)52072-9. [DOI] [PubMed] [Google Scholar]
  32. Shaw G. M., Hahn B. H., Arya S. K., Groopman J. E., Gallo R. C., Wong-Staal F. Molecular characterization of human T-cell leukemia (lymphotropic) virus type III in the acquired immune deficiency syndrome. Science. 1984 Dec 7;226(4679):1165–1171. doi: 10.1126/science.6095449. [DOI] [PubMed] [Google Scholar]
  33. Steimer K. S., Higgins K. W., Powers M. A., Stephans J. C., Gyenes A., George-Nascimento C., Luciw P. A., Barr P. J., Hallewell R. A., Sanchez-Pescador R. Recombinant polypeptide from the endonuclease region of the acquired immune deficiency syndrome retrovirus polymerase (pol) gene detects serum antibodies in most infected individuals. J Virol. 1986 Apr;58(1):9–16. doi: 10.1128/jvi.58.1.9-16.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Steimer K. S., Puma J. P., Power M. D., Powers M. A., George-Nascimento C., Stephans J. C., Levy J. A., Sanchez-Pescador R., Luciw P. A., Barr P. J. Differential antibody responses of individuals infected with AIDS-associated retroviruses surveyed using the viral core antigen p25gag expressed in bacteria. Virology. 1986 Apr 15;150(1):283–290. doi: 10.1016/0042-6822(86)90289-8. [DOI] [PubMed] [Google Scholar]
  35. Thorn R. M., Beltz G. A., Hung C. H., Fallis B. F., Winkle S., Cheng K. L., Marciani D. J. Enzyme immunoassay using a novel recombinant polypeptide to detect human immunodeficiency virus env antibody. J Clin Microbiol. 1987 Jul;25(7):1207–1212. doi: 10.1128/jcm.25.7.1207-1212.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tribe D. E., Reed D. L., Lindell P., Kenealy W. R., Ferguson B. Q., Cybulski R., Winslow D., Waselefsky D. M., Petteway S. R., Jr Antibodies reactive with human immunodeficiency virus gag-coded antigens (gag reactive only) are a major cause of enzyme-linked immunosorbent assay reactivity in a blood donor population. J Clin Microbiol. 1988 Apr;26(4):641–647. doi: 10.1128/jcm.26.4.641-647.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wang J. J., Steel S., Wisniewolski R., Wang C. Y. Detection of antibodies to human T-lymphotropic virus type III by using a synthetic peptide of 21 amino acid residues corresponding to a highly antigenic segment of gp41 envelope protein. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6159–6163. doi: 10.1073/pnas.83.16.6159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiss S. H., Goedert J. J., Sarngadharan M. G., Bodner A. J., Gallo R. C., Blattner W. A. Screening test for HTLV-III (AIDS agent) antibodies. Specificity, sensitivity, and applications. JAMA. 1985 Jan 11;253(2):221–225. [PubMed] [Google Scholar]
  39. van der Poel C. L., Reesink H. W., Tersmette T., Lelie P. N., Huisman H., Miedema F. Blood donations reactive for HIV in Western blot, but non-infective in culture and recipients of blood. Lancet. 1986 Sep 27;2(8509):752–753. doi: 10.1016/s0140-6736(86)90273-4. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES