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. 1987 Nov-Dec;60(6):575–587.

Principles of laboratory isolation and identification of the human immunodeficiency virus (HIV).

B P Griffith 1
PMCID: PMC2590393  PMID: 3324507

Abstract

Diagnosis of human immunodeficiency virus (HIV) infections has relied most frequently on detecting the presence of HIV antibodies in sera. In many situations, however, patients management can be significantly improved if the presence of HIV can be demonstrated in patients' specimens. In this review, the need and value of HIV isolation for confirming the diagnosis of HIV, for disease staging, and for monitoring the effectiveness of antiviral therapy are discussed. The steps involved in isolation of HIV, the cell systems permissive to HIV growth, and the procedures for virus identification are reviewed. Furthermore, methods available for the direct detection of HIV in patients' specimens are summarized. Although isolation of HIV is presently an elaborate procedure, as easier methods become available, it will play a large role in the management of HIV-infected individuals.

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Selected References

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

  1. Albert J., Bredberg U., Chiodi F., Böttiger B., Fenyö E. M., Norrby E., Biberfeld G. A new human retrovirus isolate of West African origin (SBL-6669) and its relationship to HTLV-IV, LAV-II, and HTLV-IIIB. AIDS Res Hum Retroviruses. 1987 Spring;3(1):3–10. doi: 10.1089/aid.1987.3.3. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Asjö B., Morfeldt-Månson L., Albert J., Biberfeld G., Karlsson A., Lidman K., Fenyö E. M. Replicative capacity of human immunodeficiency virus from patients with varying severity of HIV infection. Lancet. 1986 Sep 20;2(8508):660–662. [PubMed] [Google Scholar]
  4. 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]
  5. Chaisson R. E., Allain J. P., Leuther M., Volberding P. A. Significant changes in HIV antigen level in the serum of patients treated with azidothymidine. N Engl J Med. 1986 Dec 18;315(25):1610–1611. doi: 10.1056/NEJM198612183152511. [DOI] [PubMed] [Google Scholar]
  6. Clavel F., Mansinho K., Chamaret S., Guetard D., Favier V., Nina J., Santos-Ferreira M. O., Champalimaud J. L., Montagnier L. Human immunodeficiency virus type 2 infection associated with AIDS in West Africa. N Engl J Med. 1987 May 7;316(19):1180–1185. doi: 10.1056/NEJM198705073161903. [DOI] [PubMed] [Google Scholar]
  7. Dalgleish A. G., Beverley P. C., Clapham P. R., Crawford D. H., Greaves M. F., Weiss R. A. The CD4 (T4) antigen is an essential component of the receptor for the AIDS retrovirus. Nature. 1984 Dec 20;312(5996):763–767. doi: 10.1038/312763a0. [DOI] [PubMed] [Google Scholar]
  8. Folks T., Benn S., Rabson A., Theodore T., Hoggan M. D., Martin M., Lightfoote M., Sell K. Characterization of a continuous T-cell line susceptible to the cytopathic effects of the acquired immunodeficiency syndrome (AIDS)-associated retrovirus. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4539–4543. doi: 10.1073/pnas.82.13.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Getchell J. P., Heath J. L., Hicks D. R., Sporborg C., Mann J. M., McCormick J. B. Detection of human T cell leukemia virus type I and human immunodeficiency virus in cultured lymphocytes of a Zairian man with AIDS. J Infect Dis. 1987 Apr;155(4):612–616. doi: 10.1093/infdis/155.4.612. [DOI] [PubMed] [Google Scholar]
  11. Goudsmit J., de Wolf F., Paul D. A., Epstein L. G., Lange J. M., Krone W. J., Speelman H., Wolters E. C., Van der Noordaa J., Oleske J. M. Expression of human immunodeficiency virus antigen (HIV-Ag) in serum and cerebrospinal fluid during acute and chronic infection. Lancet. 1986 Jul 26;2(8500):177–180. doi: 10.1016/s0140-6736(86)92485-2. [DOI] [PubMed] [Google Scholar]
  12. Harada S., Koyanagi Y., Yamamoto N. Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay. Science. 1985 Aug 9;229(4713):563–566. doi: 10.1126/science.2992081. [DOI] [PubMed] [Google Scholar]
  13. Harada S., Koyanagi Y., Yamamoto N. Infection of human T-lymphotropic virus type-I (HTLV-I)-bearing MT-4 cells with HTLV-III (AIDS virus): chronological studies of early events. Virology. 1985 Oct 30;146(2):272–281. doi: 10.1016/0042-6822(85)90010-8. [DOI] [PubMed] [Google Scholar]
  14. Harnish D. G., Hammerberg O., Walker I. R., Rosenthal K. L. Early detection of HIV infection in a newborn. N Engl J Med. 1987 Jan 29;316(5):272–273. doi: 10.1056/NEJM198701293160510. [DOI] [PubMed] [Google Scholar]
  15. Harper M. E., Marselle L. M., Gallo R. C., Wong-Staal F. Detection of lymphocytes expressing human T-lymphotropic virus type III in lymph nodes and peripheral blood from infected individuals by in situ hybridization. Proc Natl Acad Sci U S A. 1986 Feb;83(3):772–776. doi: 10.1073/pnas.83.3.772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hedenskog M., Dewhurst S., Ludvigsen C., Sinangil F., Rodriguez L., Wu Y. T., Volsky D. J. Testing for antibodies to AIDS-associated retrovirus (HTLV-III/LAV) by indirect fixed cell immunofluorescence: specificity, sensitivity, and applications. J Med Virol. 1986 Aug;19(4):325–334. doi: 10.1002/jmv.1890190405. [DOI] [PubMed] [Google Scholar]
  17. Henderson L. E., Sowder R., Copeland T. D., Oroszlan S., Arthur L. O., Robey W. G., Fischinger P. J. Direct identification of class II histocompatibility DR proteins in preparations of human T-cell lymphotropic virus type III. J Virol. 1987 Feb;61(2):629–632. doi: 10.1128/jvi.61.2.629-632.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Higgins J. R., Pedersen N. C., Carlson J. R. Detection and differentiation by sandwich enzyme-linked immunosorbent assay of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus- and acquired immunodeficiency syndrome-associated retroviruslike clinical isolates. J Clin Microbiol. 1986 Sep;24(3):424–430. doi: 10.1128/jcm.24.3.424-430.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ho D. D., Rota T. R., Hirsch M. S. Infection of monocyte/macrophages by human T lymphotropic virus type III. J Clin Invest. 1986 May;77(5):1712–1715. doi: 10.1172/JCI112491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hoshino H., Shimoyama M., Miwa M., Sugimura T. Detection of lymphocytes producing a human retrovirus associated with adult T-cell leukemia by syncytia induction assay. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7337–7341. doi: 10.1073/pnas.80.23.7337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kanki P. J., M'Boup S., Ricard D., Barin F., Denis F., Boye C., Sangare L., Travers K., Albaum M., Marlink R. Human T-lymphotropic virus type 4 and the human immunodeficiency virus in West Africa. Science. 1987 May 15;236(4803):827–831. doi: 10.1126/science.3033826. [DOI] [PubMed] [Google Scholar]
  22. Kanner S. B., Parks E. S., Scott G. B., Parks W. P. Simultaneous infections with human T cell leukemia virus type I and the human immunodeficiency virus. J Infect Dis. 1987 Apr;155(4):617–625. doi: 10.1093/infdis/155.4.617. [DOI] [PubMed] [Google Scholar]
  23. Klatzmann D., Barré-Sinoussi F., Nugeyre M. T., Danquet C., Vilmer E., Griscelli C., Brun-Veziret F., Rouzioux C., Gluckman J. C., Chermann J. C. Selective tropism of lymphadenopathy associated virus (LAV) for helper-inducer T lymphocytes. Science. 1984 Jul 6;225(4657):59–63. doi: 10.1126/science.6328660. [DOI] [PubMed] [Google Scholar]
  24. Kwok S., Mack D. H., Mullis K. B., Poiesz B., Ehrlich G., Blair D., Friedman-Kien A., Sninsky J. J. Identification of human immunodeficiency virus sequences by using in vitro enzymatic amplification and oligomer cleavage detection. J Virol. 1987 May;61(5):1690–1694. doi: 10.1128/jvi.61.5.1690-1694.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lange J. M., Paul D. A., Huisman H. G., de Wolf F., van den Berg H., Coutinho R. A., Danner S. A., van der Noordaa J., Goudsmit J. Persistent HIV antigenaemia and decline of HIV core antibodies associated with transition to AIDS. Br Med J (Clin Res Ed) 1986 Dec 6;293(6560):1459–1462. doi: 10.1136/bmj.293.6560.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Levy J. A., Kaminsky L. S., Morrow W. J., Steimer K., Luciw P., Dina D., Hoxie J., Oshiro L. Infection by the retrovirus associated with the acquired immunodeficiency syndrome. Clinical, biological, and molecular features. Ann Intern Med. 1985 Nov;103(5):694–699. doi: 10.7326/0003-4819-103-5-694. [DOI] [PubMed] [Google Scholar]
  29. Levy J. A., Shimabukuro J., McHugh T., Casavant C., Stites D., Oshiro L. AIDS-associated retroviruses (ARV) can productively infect other cells besides human T helper cells. Virology. 1985 Dec;147(2):441–448. doi: 10.1016/0042-6822(85)90146-1. [DOI] [PubMed] [Google Scholar]
  30. Lifson J. D., Reyes G. R., McGrath M. S., Stein B. S., Engleman E. G. AIDS retrovirus induced cytopathology: giant cell formation and involvement of CD4 antigen. Science. 1986 May 30;232(4754):1123–1127. doi: 10.1126/science.3010463. [DOI] [PubMed] [Google Scholar]
  31. McDougal J. S., Cort S. P., Kennedy M. S., Cabridilla C. D., Feorino P. M., Francis D. P., Hicks D., Kalyanaraman V. S., Martin L. S. Immunoassay for the detection and quantitation of infectious human retrovirus, lymphadenopathy-associated virus (LAV). J Immunol Methods. 1985 Jan 21;76(1):171–183. doi: 10.1016/0022-1759(85)90489-2. [DOI] [PubMed] [Google Scholar]
  32. Monroe J. E., Andrews C., Sullivan J. L., Mulder C. Use of cytoplasmic dot-blot hybridization to detect human immunodeficiency virus RNA sequences in cultures of peripheral blood. J Infect Dis. 1987 Feb;155(2):320–322. doi: 10.1093/infdis/155.2.320. [DOI] [PubMed] [Google Scholar]
  33. Montagnier L., Gruest J., Chamaret S., Dauguet C., Axler C., Guétard D., Nugeyre M. T., Barré-Sinoussi F., Chermann J. C., Brunet J. B. Adaptation of lymphadenopathy associated virus (LAV) to replication in EBV-transformed B lymphoblastoid cell lines. Science. 1984 Jul 6;225(4657):63–66. doi: 10.1126/science.6328661. [DOI] [PubMed] [Google Scholar]
  34. Morgan D. A., Ruscetti F. W., Gallo R. Selective in vitro growth of T lymphocytes from normal human bone marrows. Science. 1976 Sep 10;193(4257):1007–1008. doi: 10.1126/science.181845. [DOI] [PubMed] [Google Scholar]
  35. Nakashima H., Koyanagi Y., Harada S., Yamamoto N. Quantitative evaluations of the effect of UV irradiation on the infectivity of HTLV-III (AIDS virus) with HTLV-I-carrying cell line, MT-4. J Invest Dermatol. 1986 Aug;87(2):239–243. doi: 10.1111/1523-1747.ep12696622. [DOI] [PubMed] [Google Scholar]
  36. Nicholson J. K., Cross G. D., Callaway C. S., McDougal J. S. In vitro infection of human monocytes with human T lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV). J Immunol. 1986 Jul 1;137(1):323–329. [PubMed] [Google Scholar]
  37. Palmer E., Sporborg C., Harrison A., Martin M. L., Feorino P. Morphology and immunoelectron microscopy of AIDS virus. Arch Virol. 1985;85(3-4):189–196. doi: 10.1007/BF01314230. [DOI] [PubMed] [Google Scholar]
  38. Pekovic D., Garzon S., Strykowski H., Ajdukovic D., Gornitsky M., Dupuy J. M. Immunogold labeling of HTLV-III/LAV in H9 cells studied by transmission electron microscopy. J Virol Methods. 1986 Jun;13(3):265–269. doi: 10.1016/0166-0934(86)90020-0. [DOI] [PubMed] [Google Scholar]
  39. Popovic M., Read-Connole E., Gallo R. C. T4 positive human neoplastic cell lines susceptible to and permissive for HTLV-III. Lancet. 1984 Dec 22;2(8417-8418):1472–1473. doi: 10.1016/s0140-6736(84)91666-0. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Ruscetti F. W., Morgan D. A., Gallo R. C. Functional and morphologic characterization of human T cells continuously grown in vitro. J Immunol. 1977 Jul;119(1):131–138. [PubMed] [Google Scholar]
  42. Rübsamen-Waigmann H., Becker W. B., Helm E. B., Brodt R., Fischer H., Henco K., Brede H. D. Isolation of variants of lymphocytopathic retroviruses from the peripheral blood and cerebrospinal fluid of patients with ARC or AIDS. J Med Virol. 1986 Aug;19(4):335–344. doi: 10.1002/jmv.1890190406. [DOI] [PubMed] [Google Scholar]
  43. Sandstrom E. G., Schooley R. T., Ho D. D., Byington R., Sarngadharan M. G., MacLane M. E., Essex M., Gallo R. C., Hirsch M. S. Detection of human anti-HTLV-III antibodies by indirect immunofluorescence using fixed cells. Transfusion. 1985 Jul-Aug;25(4):308–312. doi: 10.1046/j.1537-2995.1985.25485273806.x. [DOI] [PubMed] [Google Scholar]
  44. Tersmette M., Miedema F., Huisman H. G., Goudsmit J., Melief C. J. Productive HTLV-III infection of human B cell lines. Lancet. 1985 Apr 6;1(8432):815–816. doi: 10.1016/s0140-6736(85)91469-2. [DOI] [PubMed] [Google Scholar]
  45. Toyoshima K., Vogt P. K. Enhancement and inhibition of avian sarcoma viruses by polycations and polyanions. Virology. 1969 Jul;38(3):414–426. doi: 10.1016/0042-6822(69)90154-8. [DOI] [PubMed] [Google Scholar]

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