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. 1994 Jun;32(6):1449–1456. doi: 10.1128/jcm.32.6.1449-1456.1994

Enzyme immunoassay using native envelope glycoprotein (gp160) for detection of human immunodeficiency virus type 1 antibodies.

B C Nair 1, G Ford 1, V S Kalyanaraman 1, M Zafari 1, C Fang 1, M G Sarngadharan 1
PMCID: PMC264018  PMID: 8077388

Abstract

An enzyme immunoassay using the purified native gp160 for the detection of human immunodeficiency virus type 1 (HIV-1) antibody was developed. This assay was determined to be highly specific, since (i) 157 serum samples that were confirmed negative by Western blot (immunoblot) (WB) were negative, (ii) 41 serum samples from populations with medical conditions that might cause nonspecific assay reactivity were all negative, and (iii) all 15 serum samples that showed false-positive reactions in one or more commercial HIV-1 screening tests were negative. The assay gave 100% specificity with a randomly selected and unlinked panel of 1,000 serum samples from healthy blood donors. The sensitivity of the assay was assessed by testing 238 samples confirmed as HIV-1 antibody positive by a standardized WB assay. All 238 serum samples (100%) were reactive in the native gp160 assay. In a dilution panel of 14 weakly WB-positive serum samples, 7 samples reacted two-to fivefold more strongly in the gp160 assay than in a virus lysate-based assay; the remaining 7 samples gave comparable reactivities in the two tests. The reactivities of 13 of these 14 serum samples in the gp160 assay were higher than in a commercial enzyme immunoassay that uses a recombinant envelope protein as the antigen. The native gp160 assay was more sensitive to identify seroconversion. In a well-characterized panel of sequential blood samples from a seroconverter, the new assay detected antibodies at least one sample ahead of the other commercial assays tested.

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Selected References

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  1. Allain J. P., Laurian Y., Paul D. A., Senn D. Serological markers in early stages of human immunodeficiency virus infection in haemophiliacs. Lancet. 1986 Nov 29;2(8518):1233–1236. doi: 10.1016/s0140-6736(86)92673-5. [DOI] [PubMed] [Google Scholar]
  2. Allan J. S., Coligan J. E., Barin F., McLane M. F., Sodroski J. G., Rosen C. A., Haseltine W. A., Lee T. H., Essex M. Major glycoprotein antigens that induce antibodies in AIDS patients are encoded by HTLV-III. Science. 1985 May 31;228(4703):1091–1094. doi: 10.1126/science.2986290. [DOI] [PubMed] [Google Scholar]
  3. Anderson D. W., Epstein J. S., Lee T. H., Lairmore M. D., Saxinger C., Kalyanaraman V. S., Slamon D., Parks W., Poiesz B. J., Pierik L. T. Serological confirmation of human T-lymphotropic virus type I infection in healthy blood and plasma donors. Blood. 1989 Nov 15;74(7):2585–2591. [PubMed] [Google Scholar]
  4. Barin F., McLane M. F., Allan J. S., Lee T. H., Groopman J. E., Essex M. Virus envelope protein of HTLV-III represents major target antigen for antibodies in AIDS patients. Science. 1985 May 31;228(4703):1094–1096. doi: 10.1126/science.2986291. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Behets F., Disasi A., Ryder R. W., Bishagara K., Piot P., Kashamuka M., Kamenga M., Nzila N., Laga M., Vercauteren G. Comparison of five commercial enzyme-linked immunosorbent assays and Western immunoblotting for human immunodeficiency virus antibody detection in serum samples from Central Africa. J Clin Microbiol. 1991 Oct;29(10):2280–2284. doi: 10.1128/jcm.29.10.2280-2284.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Biggar R. J., Melbye M., Ebbesen P., Alexander S., Nielsen J. O., Sarin P., Faber V. Variation in human T lymphotropic virus III (HTLV-III) antibodies in homosexual men: decline before onset of illness related to acquired immune deficiency syndrome (AIDS). Br Med J (Clin Res Ed) 1985 Oct 12;291(6501):997–998. doi: 10.1136/bmj.291.6501.997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bolognesi D. P. Immunobiology of the human immunodeficiency virus envelope and its relationship to vaccine strategies. Mol Biol Med. 1990 Feb;7(1):1–15. [PubMed] [Google Scholar]
  9. Brun-Vezinet F., Rouzioux C., Barre-Sinoussi F., Klatzmann D., Saimot A. G., Rozenbaum W., Christol D., Gluckmann J. C., Montagnier L., Chermann J. C. Detection of IgG antibodies to lymphadenopathy-associated virus in patients with AIDS or lymphadenopathy syndrome. Lancet. 1984 Jun 9;1(8389):1253–1256. doi: 10.1016/s0140-6736(84)92444-9. [DOI] [PubMed] [Google Scholar]
  10. Burke D. S., Redfield R. R., Putman P., Alexander S. S. Variations in Western blot banding patterns of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus. J Clin Microbiol. 1987 Jan;25(1):81–84. doi: 10.1128/jcm.25.1.81-84.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Bäcker U., Gathof A., Howe J., Strobel E., Deinhardt F., Gürtler L., Grob P., Joller-Jemelka H., Kühnl P., Seidl S. A new second-generation anti-HIV-1 enzyme immunoassay using recombinant envelope and core proteins. AIDS. 1988 Dec;2(6):477–480. doi: 10.1097/00002030-198812000-00011. [DOI] [PubMed] [Google Scholar]
  12. DeVico A. L., Veronese F. D., Lee S. L., Gallo R. C., Sarngadharan M. G. High prevalence of serum antibodies to reverse transcriptase in HIV-1-infected individuals. AIDS Res Hum Retroviruses. 1988 Feb;4(1):17–22. doi: 10.1089/aid.1988.4.17. [DOI] [PubMed] [Google Scholar]
  13. Deinhardt F., Eberle J., Gürtler L. Sensitivity and specificity of eight commercial and one recombinant anti-HIV ELISA tests. Lancet. 1987 Jan 3;1(8523):40–40. doi: 10.1016/s0140-6736(87)90726-4. [DOI] [PubMed] [Google Scholar]
  14. Döpel S. H., Porstmann T., Henklein P., von Baehr R. Fine mapping of an immunodominant region of the transmembrane protein of the human immunodeficiency virus (HIV-1). J Virol Methods. 1989 Aug;25(2):167–177. doi: 10.1016/0166-0934(89)90030-x. [DOI] [PubMed] [Google Scholar]
  15. Esteban J. I., Shih J. W., Tai C. C., Bodner A. J., Kay J. W., Alter H. J. Importance of western blot analysis in predicting infectivity of anti-HTLV-III/LAV positive blood. Lancet. 1985 Nov 16;2(8464):1083–1086. doi: 10.1016/s0140-6736(85)90683-x. [DOI] [PubMed] [Google Scholar]
  16. Gallo R. C., Salahuddin S. Z., Popovic M., Shearer G. M., Kaplan M., Haynes B. F., Palker T. J., Redfield R., Oleske J., Safai B. Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Science. 1984 May 4;224(4648):500–503. doi: 10.1126/science.6200936. [DOI] [PubMed] [Google Scholar]
  17. Gnann J. W., Jr, McCormick J. B., Mitchell S., Nelson J. A., Oldstone M. B. Synthetic peptide immunoassay distinguishes HIV type 1 and HIV type 2 infections. Science. 1987 Sep 11;237(4820):1346–1349. doi: 10.1126/science.2888192. [DOI] [PubMed] [Google Scholar]
  18. Gnann J. W., Jr, Nelson J. A., Oldstone M. B. Fine mapping of an immunodominant domain in the transmembrane glycoprotein of human immunodeficiency virus. J Virol. 1987 Aug;61(8):2639–2641. doi: 10.1128/jvi.61.8.2639-2641.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gorse G. J., Frey S. E., Newman F. K., Belshe R. B. Detection of binding antibodies to native and recombinant human immunodeficiency virus type 1 envelope glycoproteins following recombinant gp160 immunization measured by flow cytometry and enzyme immunoassays. The AIDS Vaccine Clinical Trials Network. J Clin Microbiol. 1992 Oct;30(10):2606–2612. doi: 10.1128/jcm.30.10.2606-2612.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gürtler L. G., Eberle J., Lorbeer B., Deinhardt F. Sensitivity and specificity of commercial ELISA kits for screening anti-LAV/HTLV III. J Virol Methods. 1987 Jan;15(1):11–23. doi: 10.1016/0166-0934(87)90044-9. [DOI] [PubMed] [Google Scholar]
  21. Hardy C. T., Damrow T. A., Villareal D. B., Kenny G. E. Evaluation of viral-lysate enzyme-linked immunosorbent assay kits for detecting human immunodeficiency virus (type 1) infections using human sera standardized by quantitative western blotting. J Virol Methods. 1992 Jun;37(3):259–273. doi: 10.1016/0166-0934(92)90028-c. [DOI] [PubMed] [Google Scholar]
  22. Ho D. D., Sarngadharan M. G., Resnick L., Dimarzoveronese F., Rota T. R., Hirsch M. S. Primary human T-lymphotropic virus type III infection. Ann Intern Med. 1985 Dec;103(6 ):880–883. doi: 10.7326/0003-4819-103-6-880. [DOI] [PubMed] [Google Scholar]
  23. Hoxie J. A., Fitzharris T. P., Youngbar P. R., Matthews D. M., Rackowski J. L., Radka S. F. Nonrandom association of cellular antigens with HTLV-III virions. Hum Immunol. 1987 Jan;18(1):39–52. doi: 10.1016/0198-8859(87)90111-x. [DOI] [PubMed] [Google Scholar]
  24. Kalyanaraman V. S., Pal R., Gallo R. C., Sarngadharan M. G. A unique human immunodeficiency virus culture secreting soluble gp160. AIDS Res Hum Retroviruses. 1988 Oct;4(5):319–329. doi: 10.1089/aid.1988.4.319. [DOI] [PubMed] [Google Scholar]
  25. Kalyanaraman V. S., Rodriguez V., Josephs S., Gallo R. C., Sarngadharan M. G. Differential viral gene expression and its effect on the biological properties of the cell clones of an HIV-1-infected cell line. Virology. 1990 Jul;177(1):380–383. doi: 10.1016/0042-6822(90)90495-d. [DOI] [PubMed] [Google Scholar]
  26. Kalyanaraman V. S., Rodriguez V., Veronese F., Rahman R., Lusso P., DeVico A. L., Copeland T., Oroszlan S., Gallo R. C., Sarngadharan M. G. Characterization of the secreted, native gp120 and gp160 of the human immunodeficiency virus type 1. AIDS Res Hum Retroviruses. 1990 Mar;6(3):371–380. doi: 10.1089/aid.1990.6.371. [DOI] [PubMed] [Google Scholar]
  27. Kenealy W., Reed D., Cybulski R., Tribe D., Taylor P., Stevens C., Matthews T., Petteway S. Analysis of human serum antibodies to human immunodeficiency virus (HIV) using recombinant ENV and GAG antigens. AIDS Res Hum Retroviruses. 1987 Spring;3(1):95–105. doi: 10.1089/aid.1987.3.95. [DOI] [PubMed] [Google Scholar]
  28. Kitchen L., Malone G., Orgad S., Barin F., Zaizov R., Ramot B., Gazit E., Kreiss J., Leal M., Wichmann I. Viral envelope protein of HTLV-III is the major target antigen for antibodies in hemophiliac patients. J Infect Dis. 1986 Apr;153(4):788–790. doi: 10.1093/infdis/153.4.788. [DOI] [PubMed] [Google Scholar]
  29. Kühnl P., Seidl S., Holzberger G. HLA DR4 antibodies cause positive HTLV-III antibody ELISA results. Lancet. 1985 May 25;1(8439):1222–1223. doi: 10.1016/s0140-6736(85)92910-1. [DOI] [PubMed] [Google Scholar]
  30. Lasky L. A., Groopman J. E., Fennie C. W., Benz P. M., Capon D. J., Dowbenko D. J., Nakamura G. R., Nunes W. M., Renz M. E., Berman P. W. Neutralization of the AIDS retrovirus by antibodies to a recombinant envelope glycoprotein. Science. 1986 Jul 11;233(4760):209–212. doi: 10.1126/science.3014647. [DOI] [PubMed] [Google Scholar]
  31. Manca N., di Marzo Veronese F., Ho D. D., Gallo R. C., Sarngadharan M. G. Sequential changes in antibody levels to the env and gag antigens in human immunodeficiency virus infected subjects. Eur J Epidemiol. 1987 Jun;3(2):96–102. doi: 10.1007/BF00239745. [DOI] [PubMed] [Google Scholar]
  32. Moore J. P., Wallace L. A., Follett E. A., McKeating J. A. An enzyme-linked immunosorbent assay for antibodies to the envelope glycoproteins of divergent strains of HIV-1. AIDS. 1989 Mar;3(3):155–163. doi: 10.1097/00002030-198903000-00006. [DOI] [PubMed] [Google Scholar]
  33. Norrby E., Biberfeld G., Chiodi F., von Gegerfeldt A., Nauclér A., Parks E., Lerner R. Discrimination between antibodies to HIV and to related retroviruses using site-directed serology. Nature. 1987 Sep 17;329(6136):248–250. doi: 10.1038/329248a0. [DOI] [PubMed] [Google Scholar]
  34. Pal R., Nair B. C., Hoke G. M., Sarngadharan M. G., Edidin M. Lateral diffusion of CD4 on the surface of a human neoplastic T-cell line probed with a fluorescent derivative of the envelope glycoprotein (gp120) of human immunodeficiency virus type 1 (HIV-1). J Cell Physiol. 1991 May;147(2):326–332. doi: 10.1002/jcp.1041470219. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Ranki A., Valle S. L., Krohn M., Antonen J., Allain J. P., Leuther M., Franchini G., Krohn K. Long latency precedes overt seroconversion in sexually transmitted human-immunodeficiency-virus infection. Lancet. 1987 Sep 12;2(8559):589–593. doi: 10.1016/s0140-6736(87)92985-0. [DOI] [PubMed] [Google Scholar]
  37. Redfield R. R., Birx D. L., Ketter N., Tramont E., Polonis V., Davis C., Brundage J. F., Smith G., Johnson S., Fowler A. A phase I evaluation of the safety and immunogenicity of vaccination with recombinant gp160 in patients with early human immunodeficiency virus infection. Military Medical Consortium for Applied Retroviral Research. N Engl J Med. 1991 Jun 13;324(24):1677–1684. doi: 10.1056/NEJM199106133242401. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Samuel K. P., Seth A., Zweig M., Showalter S. D., Papas T. S. Bacterial expression and characterization of nine polypeptides encoded by segments of the envelope gene of human immunodeficiency virus. Gene. 1988 Apr 15;64(1):121–134. doi: 10.1016/0378-1119(88)90486-6. [DOI] [PubMed] [Google Scholar]
  40. Sarngadharan M. G., Popovic M., Bruch L., Schüpbach J., Gallo R. C. Antibodies reactive with human T-lymphotropic retroviruses (HTLV-III) in the serum of patients with AIDS. Science. 1984 May 4;224(4648):506–508. doi: 10.1126/science.6324345. [DOI] [PubMed] [Google Scholar]
  41. Schulz T. F., Aschauer J. M., Hengster P., Larcher C., Wachter H., Fleckenstein B., Dierich M. P. Envelope gene-derived recombinant peptide in the serodiagnosis of human immunodeficiency virus infection. Lancet. 1986 Jul 12;2(8498):111–112. doi: 10.1016/s0140-6736(86)91648-x. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Veronese F. D., Copeland T. D., Oroszlan S., Gallo R. C., Sarngadharan M. G. Biochemical and immunological analysis of human immunodeficiency virus gag gene products p17 and p24. J Virol. 1988 Mar;62(3):795–801. doi: 10.1128/jvi.62.3.795-801.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]
  46. Zweig M., Showalter S. D., Bladen S. V., Gilden R. V., Arthur L. O., Robey W. G., Nara P. L., Fischinger P. J. Partial purification of native HIV transmembrane protein gp41: generation of polyclonal and monoclonal antibodies. AIDS Res Hum Retroviruses. 1988 Feb;4(1):51–62. doi: 10.1089/aid.1988.4.51. [DOI] [PubMed] [Google Scholar]
  47. di Marzo Veronese F., Sarngadharan M. G., Rahman R., Markham P. D., Popovic M., Bodner A. J., Gallo R. C. Monoclonal antibodies specific for p24, the major core protein of human T-cell leukemia virus type III. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5199–5202. doi: 10.1073/pnas.82.15.5199. [DOI] [PMC free article] [PubMed] [Google Scholar]

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