title: Detection of human immunodeficiency virus type 1 (HIV-1) DNA and RNA sequences in HIV-1 antibody-positive blood donors in Uganda by the Roche AMPLICOR assay.
abstract: The ability of commercially available PCR-based assays to accurately detect or quantitate human immunodeficiency virus type 1 (HIV-1) DNA or RNA in individuals predominantly infected with HIV-1 subtypes A and D is not known. Therefore, peripheral leukocytes from 43 individuals in Kampala, Uganda, positive for HIV by the Western blot (immunoblot) assay were tested by using the Roche AMPLICOR HIV-1 assay for the detection of DNA gag sequences. Plasma from these same individuals was tested by using the Roche HIV-1 AMPLICOR MONITOR HIV-1 assay for the quantitation of HIV-1 RNA gag sequences. In addition, peripheral leukocytes were tested for HIV-1 DNA by using a lower annealing temperature or a different primer pair for the HIV-1 pol region. The proportions of individuals with detectable HIV-1 DNA and RNA gag sequences by the Roche assays were 74 and 90%, respectively. The proportions positive for HIV-1 DNA sequences by using a 50 degrees C annealing temperature or the pol primer pair were 71 and 98%, respectively. In summary, the standard Roche assay did not detect HIV-1 DNA sequences in a significant number of HIV-1-infected individuals in Uganda. However, use of a pol primer pair increased the sensitivity of the assay to 98%. The sensitivity of the Roche AMPLICOR MONITOR assay for the detection and quantitation of HIV-1 RNA sequences was significantly higher than that of the DNA-based assay, but the efficiency of the assay, and hence, the accuracy of the values obtained with RNA, is not known. Modifications to existing assays are needed to enhance the sensitivities and accuracies of these commercially available assays for use in developing countries where non-B HIV-1 subtypes predominate.
authors.affiliation: Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, Maryland 21287, USA.
authors.forename: J B
authors.initials: JB
authors.lastname: Jackson
authors.forename: E M
authors.initials: EM
authors.lastname: Piwowar
authors.forename: J
authors.initials: J
authors.lastname: Parsons
authors.forename: P
authors.initials: P
authors.lastname: Kataaha
authors.forename: G
authors.initials: G
authors.lastname: Bihibwa
authors.forename: J
authors.initials: J
authors.lastname: Onecan
authors.forename: S
authors.initials: S
authors.lastname: Kabengera
authors.forename: S D
authors.initials: SD
authors.lastname: Kennedy
authors.forename: A
authors.initials: A
authors.lastname: Butcher
mesh_terms: D004279:DNA, Viral / Q000032:analysis*; D006679:HIV Seropositivity / Q000453:epidemiology / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D017421:Sequence Analysis; D014454:Uganda / Q000453:epidemiology
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral