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. 1997 May;35(5):1118–1121. doi: 10.1128/jcm.35.5.1118-1121.1997

Detection of human immunodeficiency virus type 1 (HIV-1) antibody by western blotting and HIV-1 DNA by PCR in patients with AIDS.

J B Jackson 1, J S Parsons 1, L S Nichols 1, N Knoble 1, S Kennedy 1, E M Piwowar 1
PMCID: PMC232714  PMID: 9114392

Abstract

The human immunodeficiency virus type 1 (HIV-1) Western blotting (immunoblotting) band patterns and the sensitivity of an HIV-1 DNA PCR assay were determined by testing the blood of patients with AIDS. Plasma and cell pellets processed from the peripheral blood of 199 patients with absolute CD4 cell counts of less than 200 cells per mm3 were tested by a licensed enzyme immunoassay (EIA; Abbott HIV-1) and Western blot assay (Cambridge-Biotech) for HIV-1 antibody. The Roache HIV-1 AMPLICOR DNA PCR assay was used to test cell pellets from 125 of the 199 patients for HIV-1 gag DNA sequences. All plasma samples from these 199 sequential patients were reactive for HIV-1 antibody by EIA and were positive by Western blot assay using the criteria recommended by the Centers for Disease Control and Prevention. The majority of samples (192 of 199; 96.5%) displayed at least six of nine bands characteristic of the virus by Western blotting, with the lowest number of bands characteristic of the virus displayed by any sample being three. However, 39 and 48% of all patients exhibited no bands to p17 and p55 antigens, respectively, whereas 0 to 7.5% of all patients exhibited no bands to the other antigens. HIV-1 gag DNA sequences were detected in 117 (93.6%) of 125 cell pellets processed from the peripheral blood of these same patients. All eight patients initially negative by PCR tested positive when a second pellet which had been produced from the same blood sample was tested. Despite a decrease in antibody reactivity to HIV Gag and Pol proteins, patients with advanced HIV-1 infection remained positive for HIV-1 antibody by EIA and Western blot testing. Confirmation by the HIV-1 Western blot assay still appears to be the more sensitive assay for the diagnosis of HIV-1 infection in those individuals with advanced HIV-1 infection in the United States.

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Selected References

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  1. 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]
  2. 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]
  3. 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]
  4. Dodd R. Y., Fang C. T. The western immunoblot procedure for HIV antibodies and its interpretation. Arch Pathol Lab Med. 1990 Mar;114(3):240–245. [PubMed] [Google Scholar]
  5. Folks T. M., Powell D., Lightfoote M., Koenig S., Fauci A. S., Benn S., Rabson A., Daugherty D., Gendelman H. E., Hoggan M. D. Biological and biochemical characterization of a cloned Leu-3- cell surviving infection with the acquired immune deficiency syndrome retrovirus. J Exp Med. 1986 Jul 1;164(1):280–290. doi: 10.1084/jem.164.1.280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Jackson J. B., Drew J., Lin H. J., Otto P., Bremer J. W., Hollinger F. B., Wolinsky S. M. Establishment of a quality assurance program for human immunodeficiency virus type 1 DNA polymerase chain reaction assays by the AIDS Clinical Trials Group. ACTG PCR Working Group, and the ACTG PCR Virology Laboratories. J Clin Microbiol. 1993 Dec;31(12):3123–3128. doi: 10.1128/jcm.31.12.3123-3128.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jackson J. B., Kwok S. Y., Sninsky J. J., Hopsicker J. S., Sannerud K. J., Rhame F. S., Henry K., Simpson M., Balfour H. H., Jr Human immunodeficiency virus type 1 detected in all seropositive symptomatic and asymptomatic individuals. J Clin Microbiol. 1990 Jan;28(1):16–19. doi: 10.1128/jcm.28.1.16-19.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaminsky L. S., McHugh T., Stites D., Volberding P., Henle G., Henle W., Levy J. A. High prevalence of antibodies to acquired immune deficiency syndrome (AIDS)-associated retrovirus (ARV) in AIDS and related conditions but not in other disease states. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5535–5539. doi: 10.1073/pnas.82.16.5535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. McDougal J. S., Kennedy M. S., Nicholson J. K., Spira T. J., Jaffe H. W., Kaplan J. E., Fishbein D. B., O'Malley P., Aloisio C. H., Black C. M. Antibody response to human immunodeficiency virus in homosexual men. Relation of antibody specificity, titer, and isotype to clinical status, severity of immunodeficiency, and disease progression. J Clin Invest. 1987 Aug;80(2):316–324. doi: 10.1172/JCI113075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pan L. Z., Cheng-Mayer C., Levy J. A. Patterns of antibody response in individuals infected with the human immunodeficiency virus. J Infect Dis. 1987 Apr;155(4):626–632. doi: 10.1093/infdis/155.4.626. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Schmidt G., Amiraian K., Frey H., Stevens R. W., Berns D. S. Densitometric analysis of Western blot (immunoblot) assays for human immunodeficiency virus antibodies and correlation with clinical status. J Clin Microbiol. 1987 Oct;25(10):1993–1998. doi: 10.1128/jcm.25.10.1993-1998.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tsang V. C., Peralta J. M., Simons A. R. Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods Enzymol. 1983;92:377–391. doi: 10.1016/0076-6879(83)92032-3. [DOI] [PubMed] [Google Scholar]

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