title: Evaluation of performance of the Gen-Probe human immunodeficiency virus type 1 viral load assay using primary subtype A, C, and D isolates from Kenya.
abstract: Accurate and sensitive quantification of human immunodeficiency virus type 1 (HIV-1) RNA has been invaluable as a marker for disease prognosis and for clinical monitoring of HIV-1 disease. The first generation of commercially available HIV-1 RNA tests were optimized to detect the predominant HIV-1 subtype found in North America and Europe, subtype B. However, these tests are frequently suboptimal in detecting HIV-1 genetic forms or subtypes found in other parts of the world. The goal of the present study was to evaluate the performance of a new viral load assay with non-subtype B viruses. A transcription-mediated amplification method for detection and quantitation of diverse HIV-1 subtypes, called the Gen-Probe HIV-1 viral load assay, is under development. In this study we examined the performance of the Gen-Probe HIV-1 viral load assay relative to that of the commonly used commercial HIV-1 RNA assays using a panel of primary isolates from Kenya. For comparison, we included several subtype B cloned viruses, and we quantified each virus using an in-house quantitative-competitive reverse transcriptase PCR (QC-RT-PCR) method and gag(p24) antigen capture. The Gen-Probe HIV-1 viral load assay and a version of the Roche AMPLICOR HIV-1 MONITOR test (version 1.5) that was designed to detect a broader range of subtypes were both sensitive for the quantification of Kenyan primary isolates, which represented subtype A, C, and D viruses. The Gen-Probe HIV-1 viral load assay was more sensitive for the majority of viruses than the Roche AMPLICOR HIV-1 MONITOR test version 1.0, the Bayer Quantiplex HIV RNA 3.0 assay, or a QC-RT-PCR method in use in our laboratory, suggesting that it provides a useful method for quantifying HIV-1 RNAs from diverse parts of the world, including Africa.
authors.affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, University of Washington, Seattle, Washington 98109, USA.
authors.forename: S
authors.initials: S
authors.lastname: Emery
authors.forename: S
authors.initials: S
authors.lastname: Bodrug
authors.forename: B A
authors.initials: BA
authors.lastname: Richardson
authors.forename: C
authors.initials: C
authors.lastname: Giachetti
authors.forename: M A
authors.initials: MA
authors.lastname: Bott
authors.forename: D
authors.initials: D
authors.lastname: Panteleeff
authors.forename: L L
authors.initials: LL
authors.lastname: Jagodzinski
authors.forename: N L
authors.initials: NL
authors.lastname: Michael
authors.forename: R
authors.initials: R
authors.lastname: Nduati
authors.forename: J
authors.initials: J
authors.lastname: Bwayo
authors.forename: J K
authors.initials: JK
authors.lastname: Kreiss
authors.forename: J
authors.initials: J
authors.lastname: Overbaugh
mesh_terms: D000328:Adult; D005069:Evaluation Studies as Topic; D005260:Female; D016655:HIV Core Protein p24 / Q000097:blood; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D007231:Infant, Newborn; D007630:Kenya; D015341:Nucleic Acid Probes; D012367:RNA, Viral / Q000097:blood; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D019562:Viral Load
chemical_list: D016655:HIV Core Protein p24; D015341:Nucleic Acid Probes; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic