title: PCR-Based assay to quantify human immunodeficiency virus type 1 DNA in peripheral blood mononuclear cells.
abstract: An assay that quantifies the amount of human immunodeficiency virus type 1 (HIV-1) DNA in peripheral blood mononuclear cells has been developed. PCR amplification of the HIV-1 DNA is performed in the presence of an internal quantitation standard, and colorimetric detection of the amplified product is performed with microwell plates. The copies of HIV-1 DNA are normalized to total genomic DNA input. The assay has an analytical sensitivity of 10 input copies per amplification reaction and a three-log detection range. In an analysis of sequential samples from patients on combination therapy, HIV-1 DNA was quantifiable for all individuals tested, including those with undetectable plasma HIV-1 RNA. In a separate study, a comparison of HIV-1 DNA levels was made with a group of long-term survivors and progressors. The mean HIV-1 DNA levels were lower in the long-term survivors than in the progressors (P, 0.04). The mean HIV-1 RNA levels were also lower, but the difference was not statistically significant (P, 0.164). A quantitative DNA assay will provide an additional tool to gain insight into the natural history of infection and the continued efficacy of potent antiretroviral therapies.
authors.affiliation: Roche Molecular Systems, Inc., Alameda, California 94501, USA. Cindy.Christopherson@Roche.com
authors.forename: C
authors.initials: C
authors.lastname: Christopherson
authors.forename: Y
authors.initials: Y
authors.lastname: Kidane
authors.forename: B
authors.initials: B
authors.lastname: Conway
authors.forename: J
authors.initials: J
authors.lastname: Krowka
authors.forename: H
authors.initials: H
authors.lastname: Sheppard
authors.forename: S
authors.initials: S
authors.lastname: Kwok
mesh_terms: D019380:Anti-HIV Agents / Q000627:therapeutic use; D004279:DNA, Viral / Q000097:blood* / Q000302:isolation & purification; D005456:Fluorescent Dyes / Q000378:metabolism; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D020276:HIV Long-Term Survivors; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007958:Leukocyte Count; D007963:Leukocytes, Mononuclear / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D019380:Anti-HIV Agents; D004279:DNA, Viral; D005456:Fluorescent Dyes; D012367:RNA, Viral