Abstract
To evaluate the performance of a serological test for human immunodeficiency virus type 1 (HIV-1) infections based on the use of a recombinant envelope gene-derived protein as the antigen, we caused expression of a 1.4-kilobase fragment of HIV.DNA that codes for the complete gp41 transmembrane protein in an Escherichia coli expression vector and used Western blots (WB; immunoblots) prepared with recombinant material (pEX-41) to detect antibodies to HIV-1. This test detected all 339 sera which were positive by a combination of conventional serodiagnostic assays and produced no false-positive results with 311 negative samples. Also no false-positive results were obtained with 20 sera from systemic lupus erythematosus patients which had high titers of cross-reactive autoantibodies. In six cases, the pEX-41 WB proved to be more sensitive than individual assays applied on their own, and in five cases it was even more sensitive than a combination of conventional assays. We tested 221 sera in both our pEX-41 WB and a commercially available recombinant enzyme immunoassay (EIA [Abbott]). The results were identical in 188 cases. A total of 27 sera containing antibodies to gp41 as demonstrated in the pEX-41 WB, as well as the Abbott recombinant EIA, had no antibodies to the recombinant core antigen as measured in the Abbott EIA. However, 25 of these sera did stain the 24-kilodalton band on a WB with purified virus. Six sera that were positive in all of the conventional confirmatory assays and reacted strongly with the pEX-41 WB did not recognize the surface antigen used in the Abbott recombinant EIA. We conclude that the use of WB prepared with recombinant-derived p41 offers a very sensitive and specific method to detect antibodies to HIV.
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- 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]
- Biberfeld G., Böttiger B., Bredberg-Rådén U., Putkonen P. O., Ericsson L., Berglund O., Starup C., Hákånsson C. Findings in four HTLV-IV seropositive women from West Africa. Lancet. 1986 Dec 6;2(8519):1330–1331. doi: 10.1016/s0140-6736(86)91451-0. [DOI] [PubMed] [Google Scholar]
- Brücker G., Brun-Vezinet F., Rosenheim M., Rey M. A., Katlama C., Gentilini M. HIV-2 infection in two homosexual men in France. Lancet. 1987 Jan 24;1(8526):223–223. doi: 10.1016/s0140-6736(87)90043-2. [DOI] [PubMed] [Google Scholar]
- Carlson J. R., Mertens S. C., Yee J. L., Gardner M. B., Watson-Williams E. J., Ghrayeb J., Jennings M. B., Biggar R. J. Rapid, easy, and economical screening test for antibodies to human immunodeficiency virus. Lancet. 1987 Feb 14;1(8529):361–362. doi: 10.1016/s0140-6736(87)91730-2. [DOI] [PubMed] [Google Scholar]
- Chang N. T., Chanda P. K., Barone A. D., McKinney S., Rhodes D. P., Tam S. H., Shearman C. W., Huang J., Chang T. W., Gallo R. C. Expression in Escherichia coli of open reading frame gene segments of HTLV-III. Science. 1985 Apr 5;228(4695):93–96. doi: 10.1126/science.2983429. [DOI] [PubMed] [Google Scholar]
- Chang N. T., Huang J., Ghrayeb J., McKinney S., Chanda P. K., Chang T. W., Putney S., Sarngadharan M. G., Wong-Staal F., Gallo R. C. An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS. Nature. 1985 May 9;315(6015):151–154. doi: 10.1038/315151a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Crowl R., Ganguly K., Gordon M., Conroy R., Schaber M., Kramer R., Shaw G., Wong-Staal F., Reddy E. P. HTLV-III env gene products synthesized in E. coli are recognized by antibodies present in the sera of AIDS patients. Cell. 1985 Jul;41(3):979–986. doi: 10.1016/s0092-8674(85)80078-7. [DOI] [PubMed] [Google Scholar]
- 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]
- Goedert J. J. Testing for human immunodeficiency virus. Ann Intern Med. 1986 Oct;105(4):609–610. doi: 10.7326/0003-4819-105-4-609. [DOI] [PubMed] [Google Scholar]
- Hengster P., Larcher C., Pöckl E., Schulz T., Wachter H., Dierich M. P. Nachweis von Antikörpern gegen HTLV-III: Vergleich verschiedener ELISAs als Suchtests und des Western Blot mit der Immunfluoreszenz. Bestätigungsverfahren. Wien Klin Wochenschr. 1987 Feb 20;99(4):112–114. [PubMed] [Google Scholar]
- Lange J. M., Coutinho R. A., Krone W. J., Verdonck L. F., Danner S. A., van der Noordaa J., Goudsmit J. Distinct IgG recognition patterns during progression of subclinical and clinical infection with lymphadenopathy associated virus/human T lymphotropic virus. Br Med J (Clin Res Ed) 1986 Jan 25;292(6515):228–230. doi: 10.1136/bmj.292.6515.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mortimer P. P., Parry J. V., Mortimer J. Y. Which anti-HTLV III/LAV assays for screening and confirmatory testing? Lancet. 1985 Oct 19;2(8460):873–877. doi: 10.1016/s0140-6736(85)90136-9. [DOI] [PubMed] [Google Scholar]
- Mølbak K., Lauritzen E., Fernandes D., Böttiger B., Biberfeld G. Antibodies to HTLV-IV associated with chronic, fatal illness resembling "slim" disease. Lancet. 1986 Nov 22;2(8517):1214–1215. doi: 10.1016/s0140-6736(86)92217-8. [DOI] [PubMed] [Google Scholar]
- 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]
- Ratner L., Haseltine W., Patarca R., Livak K. J., Starcich B., Josephs S. F., Doran E. R., Rafalski J. A., Whitehorn E. A., Baumeister K. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature. 1985 Jan 24;313(6000):277–284. doi: 10.1038/313277a0. [DOI] [PubMed] [Google Scholar]
- Salahuddin S. Z., Groopman J. E., Markham P. D., Sarngadharan M. G., Redfield R. R., McLane M. F., Essex M., Sliski A., Gallo R. C. HTLV-III in symptom-free seronegative persons. Lancet. 1984 Dec 22;2(8417-8418):1418–1420. doi: 10.1016/s0140-6736(84)91619-2. [DOI] [PubMed] [Google Scholar]
- 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]
- Schneider J., Bayer H., Bienzle U., Hunsmann G. A glycopolypeptide (gp 100) is the main antigen detected by HTLV-III antisera. Med Microbiol Immunol. 1985;174(1):35–42. doi: 10.1007/BF02123669. [DOI] [PubMed] [Google Scholar]
- 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]
- Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thorpe R., Bird C., Garrett A. J., Minor P. D., Schild G. C., Thomas D. P. False-positive immunoblot results with antibodies against human immunodeficiency virus. Lancet. 1986 Sep 13;2(8507):627–628. doi: 10.1016/s0140-6736(86)92449-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Veronese F. D., DeVico A. L., Copeland T. D., Oroszlan S., Gallo R. C., Sarngadharan M. G. Characterization of gp41 as the transmembrane protein coded by the HTLV-III/LAV envelope gene. Science. 1985 Sep 27;229(4720):1402–1405. doi: 10.1126/science.2994223. [DOI] [PubMed] [Google Scholar]