Quantitation of human immunodeficiency virus type 1 DNA and RNA by a novel internally controlled PCR assay (original) (raw)

Abstract

A novel internally controlled PCR (ICPCR) assay was developed to accurately quantitate human immunodeficiency virus type 1 (HIV-1) DNA and RNA in peripheral blood mononuclear cells and plasma. The ICPCR assay was sensitive and reproducible within a linear range of amplification of 10(0) to 10(3) copies for HIV-1 DNA and 10(1) to 10(4) copies for HIV-1 RNA. The assay detected HIV-1 RNA in plasma and peripheral blood mononuclear cells from all HIV-1 subjects regardless of disease stage. ICPCR was compared with a branched-DNA signal amplification assay for subjects beginning antiretroviral therapy. The reductions in plasma HIV-1 RNA in response to therapy were comparable with the two assays. The ICPCR assay should be useful in monitoring HIV-1 RNA levels both in natural history studies and in clinical trials of antiretroviral agents.

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Selected References

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  1. Bagnarelli P., Menzo S., Valenza A., Manzin A., Giacca M., Ancarani F., Scalise G., Varaldo P. E., 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. doi: 10.1128/jvi.66.12.7328-7335.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bagnarelli P., Valenza A., Menzo S., Manzin A., Scalise G., Varaldo P. E., Clementi M. Dynamics of molecular parameters of human immunodeficiency virus type 1 activity in vivo. J Virol. 1994 Apr;68(4):2495–2502. doi: 10.1128/jvi.68.4.2495-2502.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Connor R. I., Mohri H., Cao Y., Ho D. D. Increased viral burden and cytopathicity correlate temporally with CD4+ T-lymphocyte decline and clinical progression in human immunodeficiency virus type 1-infected individuals. J Virol. 1993 Apr;67(4):1772–1777. doi: 10.1128/jvi.67.4.1772-1777.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Furtado M. R., Murphy R., Wolinsky S. M. Quantification of human immunodeficiency virus type 1 tat mRNA as a marker for assessing the efficacy of antiretroviral therapy. J Infect Dis. 1993 Jan;167(1):213–216. doi: 10.1093/infdis/167.1.213. [DOI] [PubMed] [Google Scholar]
  5. Gupta P., Kingsley L., Armstrong J., Ding M., Cottrill M., Rinaldo C. Enhanced expression of human immunodeficiency virus type 1 correlates with development of AIDS. Virology. 1993 Oct;196(2):586–595. doi: 10.1006/viro.1993.1514. [DOI] [PubMed] [Google Scholar]
  6. Holodniy M., Katzenstein D. A., Sengupta S., Wang A. M., Casipit C., Schwartz D. H., Konrad M., Groves E., Merigan T. C. Detection and quantification of human immunodeficiency virus RNA in patient serum by use of the polymerase chain reaction. J Infect Dis. 1991 Apr;163(4):862–866. doi: 10.1093/infdis/163.4.862. [DOI] [PubMed] [Google Scholar]
  7. Michael N. L., Vahey M., Burke D. S., Redfield R. R. 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. doi: 10.1128/jvi.66.1.310-316.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Montoya J. G., Wood R., Katzenstein D., Holodny M., Merigan T. C. Peripheral blood mononuclear cell human immunodeficiency virus type 1 proviral DNA quantification by polymerase chain reaction: relationship to immunodeficiency and drug effect. J Clin Microbiol. 1993 Oct;31(10):2692–2696. doi: 10.1128/jcm.31.10.2692-2696.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mulder J., McKinney N., Christopherson C., Sninsky J., Greenfield L., Kwok S. Rapid and simple PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma: application to acute retroviral infection. J Clin Microbiol. 1994 Feb;32(2):292–300. doi: 10.1128/jcm.32.2.292-300.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Piatak M., Jr, Saag M. S., Yang L. C., Clark S. J., Kappes J. C., Luk K. C., Hahn B. H., Shaw G. M., Lifson J. D. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science. 1993 Mar 19;259(5102):1749–1754. doi: 10.1126/science.8096089. [DOI] [PubMed] [Google Scholar]
  11. Schnittman S. M., Greenhouse J. J., Psallidopoulos M. C., Baseler M., Salzman N. P., Fauci A. S., Lane H. C. Increasing viral burden in CD4+ T cells from patients with human immunodeficiency virus (HIV) infection reflects rapidly progressive immunosuppression and clinical disease. Ann Intern Med. 1990 Sep 15;113(6):438–443. doi: 10.7326/0003-4819-113-6-438. [DOI] [PubMed] [Google Scholar]
  12. Wolinsky S. M., Rinaldo C. R., Kwok S., Sninsky J. J., Gupta P., Imagawa D., Farzadegan H., Jacobson L. P., Grovit K. S., Lee M. H. Human immunodeficiency virus type 1 (HIV-1) infection a median of 18 months before a diagnostic western blot. Evidence from a cohort of homosexual men. Ann Intern Med. 1989 Dec 15;111(12):961–972. doi: 10.7326/0003-4819-111-12-961. [DOI] [PubMed] [Google Scholar]