title: HIV-1 low copy viral sequencing-A prototype assay.
abstract: In HIV-1 patients with low viral burden, sequencing is often problematic, yet important. This study presents a sensitive, sub-type independent system for sequencing of low level viremia. Sequencing data from 32 HIV-1 infected patients with low level viremia were collected longitudinally. A combination of ViroSeq® HIV-1 Genotyping System and an in-house nesting protocol was used. Eight sub-types were represented. The success-rate of amplification of both PR and RT in the same sample was 100% in samples with viral loads above 100 copies/ml. Below 100 copies/ml, this study managed to amplify both regions in 7/13 (54%) samples. The assays were able to amplify either PR or RT in all sub-types included but one sub-type A specimen. In conclusion, this study presents a promising, simple assay to increase the ability to perform HIV-1 resistance testing at low level viremia. This is a prototype assay and the method needs further testing to evaluate clinical performance.
authors.affiliation: a Department of Infectious Diseases , University of Gothenburg , Sweden ;
authors.forename: Tomas
authors.initials: T
authors.lastname: Mellberg
authors.affiliation: b Department of Clinical Virology , University of Gothenburg , Sweden.
authors.forename: Jon
authors.initials: J
authors.lastname: Krabbe
authors.affiliation: a Department of Infectious Diseases , University of Gothenburg , Sweden ;
authors.forename: Magnus
authors.initials: M
authors.lastname: Gisslén
authors.affiliation: b Department of Clinical Virology , University of Gothenburg , Sweden.
authors.forename: Bo
authors.initials: B
authors.lastname: Svennerholm
mesh_terms: D001483:Base Sequence; D024882:Drug Resistance, Viral; D060005:Genotyping Techniques / Q000379:methods; D015658:HIV Infections / Q000175:diagnosis / Q000821:virology*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D009154:Mutation; D012367:RNA, Viral / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D019562:Viral Load; D014766:Viremia / Q000821:virology
chemical_list: D012367:RNA, Viral
keywords: HIV-1; low level viremia; sequencing