title: Poor sensitivity, specificity, and reproducibility of detection of HIV-1 DNA in serum by polymerase chain reaction. The Transfusion Safety Study Group.
abstract: A series of recent studies have reported detection by the polymerase chain reaction (PCR) of cell-free human immunodeficiency virus type 1 (HIV-1) DNA (as opposed to virion RNA) in serum from both seropositive and seronegative persons. To evaluate the sensitivity, specificity, and reproducibility of PCR detection of cell-free HIV-1 DNA, we distributed coded panels containing 98 serum specimens obtained from well-characterized, infected individuals and control blood donors to the two laboratories with reported experience with this technique. Positive results were reported with HIV-1 gag primers (SK38/39) for 48 of 188 separate PCR determinations on DNA extracts from 44 serum samples from seropositive patients (25.5% sensitivity). HIV-1 gag signal was also reported for 28 of 151 PCR determinations on 34 samples from noninfected blood donors (18.5% false-positive rate). PCR for HIV-1 env DNA performed in one laboratory was negative on all specimens from seropositive and seronegative patients. Results for cell-free HIV-1 gag and human genomic (beta-globin or HLA DQ-alpha) DNA were inconsistent on replicate and serial specimens evaluated within each laboratory and between laboratories. These results indicate that current techniques for detecting cell-free HIV-1 DNA in serum lack adequate sensitivity, specificity, and reproducibility for widespread clinical applications.
authors.affiliation: Irwin Memorial Blood Centers, University of California, San Francisco.
authors.forename: M P
authors.initials: MP
authors.lastname: Busch
authors.forename: D R
authors.initials: DR
authors.lastname: Henrard
authors.forename: I K
authors.initials: IK
authors.lastname: Hewlett
authors.forename: W F
authors.initials: WF
authors.lastname: Mehaffey
authors.forename: J S
authors.initials: JS
authors.lastname: Epstein
authors.forename: J P
authors.initials: JP
authors.lastname: Allain
authors.forename: T H
authors.initials: TH
authors.lastname: Lee
authors.forename: J W
authors.initials: JW
authors.lastname: Mosley
mesh_terms: D001483:Base Sequence; D004279:DNA, Viral / Q000097:blood*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral