PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
 
J Clin Microbiol. 1995 January; 33(1): 16–23.
PMCID: PMC227871

Quantitative molecular monitoring of human immunodeficiency virus type 1 activity during therapy with specific antiretroviral compounds.

Abstract

Methods for the absolute quantitation of nucleic acids present in small amounts in biological samples (competitive PCR and competitive reverse transcription PCR) were applied to the direct monitoring of specific anti-human immunodeficiency virus type 1 (HIV-1) therapy. With these techniques, different parameters of HIV-1 activity (including genomic RNA copy numbers in plasma, proviral and late transcript copy numbers in peripheral blood lymphocytes, and mean transcriptional activity per each HIV-1 provirus) were monitored during therapy with azidothymidine or ddI. In most of these treated patients, a direct response to the antiretroviral compounds employed was detected during the first few weeks of treatment, as documented by a fast decrease of all molecular indexes of HIV-1 activity. However, residual viral replication (albeit at minimal levels) was documented during therapy in all subjects monitored in this study. In a minority of the patients under study (3 of 12), the drug-dependent viral inhibition was maintained throughout the observation time (213 to 791 days), but in 9 patients a rebound in viremia level was detected during therapy with competitive reverse transcription PCR. Sequencing analysis of a portion of the HIV-1 gene pol from cell-free virions showed that circulating viral variants bearing at least two mutations compatible with azidothymidine or ddI resistance were detectable in the patients who exhibited a rebound in cell-free HIV-1 genomic RNA copy numbers in plasma but not in one patient who maintained (for 455 days) lowered levels of viral load during ddI treatment.

Full Text

The Full Text of this article is available as a PDF (227K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bagnarelli P, Menzo S, Manzin A, Giacca M, Varaldo PE, Clementi M. Detection of human immunodeficiency virus type 1 genomic RNA in plasma samples by reverse-transcription polymerase chain reaction. J Med Virol. 1991 Jun;34(2):89–95. [PubMed]
  • Bagnarelli P, Menzo S, Valenza A, Manzin A, Giacca M, Ancarani F, Scalise G, Varaldo PE, Clementi M. Molecular profile of human immunodeficiency virus type 1 infection in symptomless patients and in patients with AIDS. J Virol. 1992 Dec;66(12):7328–7335. [PMC free article] [PubMed]
  • Bagnarelli P, Valenza A, Menzo S, Manzin A, Scalise G, Varaldo PE, Clementi M. Dynamics of molecular parameters of human immunodeficiency virus type 1 activity in vivo. J Virol. 1994 Apr;68(4):2495–2502. [PMC free article] [PubMed]
  • Boeri E, Giri A, Lillo F, Ferrari G, Varnier OE, Ferro A, Sabbatani S, Saxinger WC, Franchini G. In vivo genetic variability of the human immunodeficiency virus type 2 V3 region. J Virol. 1992 Jul;66(7):4546–4550. [PMC free article] [PubMed]
  • Burns DP, Desrosiers RC. Selection of genetic variants of simian immunodeficiency virus in persistently infected rhesus monkeys. J Virol. 1991 Apr;65(4):1843–1854. [PMC free article] [PubMed]
  • Clementi M, Bagnarelli P, Manzin A, Menzo S. Competitive polymerase chain reaction and analysis of viral activity at the molecular level. Genet Anal Tech Appl. 1994;11(1):1–6. [PubMed]
  • Clementi M, Bagnarelli P, Menzo S, Valenza A, Manzin A, Varaldo PE. Clearance of HIV-1 viraemia after seroconversion. Lancet. 1993 Jan 30;341(8840):315–316. [PubMed]
  • Clementi M, Menzo S, Bagnarelli P, Manzin A, Valenza A, Varaldo PE. Quantitative PCR and RT-PCR in virology. PCR Methods Appl. 1993 Feb;2(3):191–196. [PubMed]
  • Fisher AG, Ensoli B, Looney D, Rose A, Gallo RC, Saag MS, Shaw GM, Hahn BH, Wong-Staal F. Biologically diverse molecular variants within a single HIV-1 isolate. Nature. 1988 Aug 4;334(6181):444–447. [PubMed]
  • Gilliland G, Perrin S, Blanchard K, Bunn HF. Analysis of cytokine mRNA and DNA: detection and quantitation by competitive polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2725–2729. [PubMed]
  • Kellam P, Boucher CA, Larder BA. Fifth mutation in human immunodeficiency virus type 1 reverse transcriptase contributes to the development of high-level resistance to zidovudine. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1934–1938. [PubMed]
  • Kellogg DE, Sninsky JJ, Kwok S. Quantitation of HIV-1 proviral DNA relative to cellular DNA by the polymerase chain reaction. Anal Biochem. 1990 Sep;189(2):202–208. [PubMed]
  • Koken SE, van Wamel JL, Goudsmit J, Berkhout B, Geelen JL. Natural variants of the HIV-1 long terminal repeat: analysis of promoters with duplicated DNA regulatory motifs. Virology. 1992 Dec;191(2):968–972. [PubMed]
  • Kusumi K, Conway B, Cunningham S, Berson A, Evans C, Iversen AK, Colvin D, Gallo MV, Coutre S, Shpaer EG, et al. Human immunodeficiency virus type 1 envelope gene structure and diversity in vivo and after cocultivation in vitro. J Virol. 1992 Feb;66(2):875–885. [PMC free article] [PubMed]
  • Larder BA, Kemp SD. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science. 1989 Dec 1;246(4934):1155–1158. [PubMed]
  • Manzin A, Menzo S, Bagnarelli P, Varaldo PE, Bearzi I, Carloni G, Galibert F, Clementi M. Sequence analysis of the hepatitis B virus pre-C region in hepatocellular carcinoma [HCC] and nontumoral liver tissues from HCC patients. Virology. 1992 Jun;188(2):890–895. [PubMed]
  • Menzo S, Bagnarelli P, Giacca M, Manzin A, Varaldo PE, Clementi M. Absolute quantitation of viremia in human immunodeficiency virus infection by competitive reverse transcription and polymerase chain reaction. J Clin Microbiol. 1992 Jul;30(7):1752–1757. [PMC free article] [PubMed]
  • Michael NL, Vahey M, Burke DS, Redfield RR. Viral DNA and mRNA expression correlate with the stage of human immunodeficiency virus (HIV) type 1 infection in humans: evidence for viral replication in all stages of HIV disease. J Virol. 1992 Jan;66(1):310–316. [PMC free article] [PubMed]
  • Oka S, Urayama K, Hirabayashi Y, Ohnishi K, Goto H, Mitamura K, Kimura S, Shimada K. Quantitative analysis of human immunodeficiency virus type-1 DNA in asymptomatic carriers using the polymerase chain reaction. Biochem Biophys Res Commun. 1990 Feb 28;167(1):1–8. [PubMed]
  • Pedroza Martins L, Chenciner N, Wain-Hobson S. Complex intrapatient sequence variation in the V1 and V2 hypervariable regions of the HIV-1 gp 120 envelope sequence. Virology. 1992 Dec;191(2):837–845. [PubMed]
  • Piatak M, Jr, Luk KC, Williams B, Lifson JD. Quantitative competitive polymerase chain reaction for accurate quantitation of HIV DNA and RNA species. Biotechniques. 1993 Jan;14(1):70–81. [PubMed]
  • Piatak M, Jr, Saag MS, Yang LC, Clark SJ, Kappes JC, Luk KC, Hahn BH, Shaw GM, Lifson JD. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science. 1993 Mar 19;259(5102):1749–1754. [PubMed]
  • Sanchez-Pescador R, Power MD, Barr PJ, Steimer KS, Stempien MM, Brown-Shimer SL, Gee WW, Renard A, Randolph A, Levy JA, et al. Nucleotide sequence and expression of an AIDS-associated retrovirus (ARV-2). Science. 1985 Feb 1;227(4686):484–492. [PubMed]
  • Scadden DT, Wang Z, Groopman JE. Quantitation of plasma human immunodeficiency virus type 1 RNA by competitive polymerase chain reaction. J Infect Dis. 1992 Jun;165(6):1119–1123. [PubMed]
  • Semple M, Loveday C, Weller I, Tedder R. Direct measurement of viraemia in patients infected with HIV-1 and its relationship to disease progression and zidovudine therapy. J Med Virol. 1991 Sep;35(1):38–45. [PubMed]
  • Simmonds P, Balfe P, Peutherer JF, Ludlam CA, Bishop JO, Brown AJ. Human immunodeficiency virus-infected individuals contain provirus in small numbers of peripheral mononuclear cells and at low copy numbers. J Virol. 1990 Feb;64(2):864–872. [PMC free article] [PubMed]
  • St Clair MH, Martin JL, Tudor-Williams G, Bach MC, Vavro CL, King DM, Kellam P, Kemp SD, Larder BA. Resistance to ddI and sensitivity to AZT induced by a mutation in HIV-1 reverse transcriptase. Science. 1991 Sep 27;253(5027):1557–1559. [PubMed]
  • Stieger M, Démollière C, Ahlborn-Laake L, Mous J. Competitive polymerase chain reaction assay for quantitation of HIV-1 DNA and RNA. J Virol Methods. 1991 Sep-Oct;34(2):149–160. [PubMed]
  • Yerly S, Chamot E, Hirschel B, Perrin LH. Quantitation of human immunodeficiency virus provirus and circulating virus: relationship with immunologic parameters. J Infect Dis. 1992 Aug;166(2):269–276. [PubMed]
  • Zhang LQ, Simmonds P, Ludlam CA, Brown AJ. Detection, quantification and sequencing of HIV-1 from the plasma of seropositive individuals and from factor VIII concentrates. AIDS. 1991 Jun;5(6):675–681. [PubMed]

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