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An improved PCR method for detection of HIV-1 proviral DNA of a wide range of subtypes and recombinant forms circulating globally.

Abstract
Proviral DNAs are being measured increasingly as a marker of the efficacy of highly active anti-retroviral therapy (HAART) and is accepted for the early diagnosis of perinatal HIV-1 infections. This requires a standardized test which enables the detection of a wide range of subtypes worldwide including O, N and circulating recombinant forms (CRFs). Based on a previous publication, a PCR - Test for HIV-1 provirus detection in peripheral blood mononuclear cells (PBMCs) was developed. Blood samples from 80 individuals infected with HIV-1 and 20 persons negative for HIV-1&2 from Africa and Germany were tested for the presence of HIV-1 provirus DNA. The primer system used enables the detection of proviral DNA despite the high concentrations of human DNA. The limit of detection was determined to be 5 copies per 10(5) cells. All 20 samples from persons negative for HIV were negative for HIV-1 proviral DNA while provirus DNA was amplified from 76 of the 80 (95%) samples from persons infected with HIV. The amplified products were detected by gel-electrophoresis, flow cytometry and real-time PCR. All three detection systems provided the same results.
AuthorsJürgen Weidner, Uwe Cassens, Wolfgang Göhde, Walter Sibrowski, Georgina Odaibo, David Olaleye, Doris Reichelt, Burkhard Greve
JournalJournal of virological methods (J Virol Methods) Vol. 172 Issue 1-2 Pg. 22-6 (Mar 2011) ISSN: 1879-0984 [Electronic] Netherlands
PMID21182872 (Publication Type: Journal Article)
CopyrightCopyright © 2010 Elsevier B.V. All rights reserved.
Topics
  • Electrophoresis, Agar Gel
  • Flow Cytometry
  • HIV Infections (diagnosis)
  • HIV Seropositivity
  • HIV-1 (genetics)
  • Humans
  • Leukocytes, Mononuclear (virology)
  • Polymerase Chain Reaction
  • Proviruses (genetics)
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Virology (methods)

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