title: 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.
authors.affiliation: Partec GmbH, Am Flughafen 13, 02828 Görlitz, Germany. weidnej@uni-muenster.de
authors.forename: Jürgen
authors.initials: J
authors.lastname: Weidner
authors.forename: Uwe
authors.initials: U
authors.lastname: Cassens
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Göhde
authors.forename: Walter
authors.initials: W
authors.lastname: Sibrowski
authors.forename: Georgina
authors.initials: G
authors.lastname: Odaibo
authors.forename: David
authors.initials: D
authors.lastname: Olaleye
authors.forename: Doris
authors.initials: D
authors.lastname: Reichelt
authors.forename: Burkhard
authors.initials: B
authors.lastname: Greve
mesh_terms: D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D015658:HIV Infections / Q000175:diagnosis*; D006679:HIV Seropositivity; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*