title: Multicenter evaluation of methods to quantitate human immunodeficiency virus type 1 RNA in seminal plasma.
abstract: We have evaluated two commercially available kits (AMPLICOR MONITOR [Roche] and NASBA HIV-1 QT or NucliSens HIV-1 QT [Organon Teknika]) and two noncommercial methods for the accurate quantitation of human immunodeficiency virus type 1 (HIV-1) RNA in seminal plasma. The same panels of coded specimens were tested on four separate occasions. Laboratories using the commercial assays employed silica beads to isolate HIV-1 RNA, which removed inhibitory factors sometimes found in seminal plasma. Sensitivities and specificities, respectively, for each assay were as follows: AMPLICOR MONITOR, 100 and 73%; NASBA HIV-1 QT, 84 and 100%; NucliSens HIV-1 QT, 99 and 98%; and noncommercial assays, 91 and 73%. When results from the laboratory that was inexperienced with the silica bead extraction method were excluded from the analysis, specificity for the Roche assay increased to 100%. The commercial assays demonstrated highly reproducible results, with intra-assay standard deviations (measured in log(10) RNA copies/milliliter of seminal plasma) ranging from 0.11 to 0.32; those of the noncommercial assays ranged from 0.12 to 0.75. Differences in mean estimated HIV-1 RNA concentrations were </=0.67 log(10) and were greater at low viral loads. Suspension matrices that used blood plasma or seminal plasma did not make a difference in recovery of HIV-1 RNA, which suggested that blood plasma specimens can be used as external controls for seminal plasma assays. More variation in the HIV-1 RNA viral loads was observed in the seminal plasma values than in the blood plasma values when paired specimens from HIV-1-infected men were tested. Quantitation of HIV-1 RNA in seminal plasma can be reliably accomplished using two commercially available assays, and may be incorporated into the evaluations of HIV-1 seropositive men enrolled in clinical studies.
authors.affiliation: University of North Carolina, Chapel Hill, North Carolina 27599, USA. fiscussa@med.unc.edu
authors.forename: S A
authors.initials: SA
authors.lastname: Fiscus
authors.forename: D
authors.initials: D
authors.lastname: Brambilla
authors.forename: R W
authors.initials: RW
authors.lastname: Coombs
authors.forename: B
authors.initials: B
authors.lastname: Yen-Lieberman
authors.forename: J
authors.initials: J
authors.lastname: Bremer
authors.forename: A
authors.initials: A
authors.lastname: Kovacs
authors.forename: S
authors.initials: S
authors.lastname: Rasheed
authors.forename: M
authors.initials: M
authors.lastname: Vahey
authors.forename: T
authors.initials: T
authors.lastname: Schutzbank
authors.forename: P S
authors.initials: PS
authors.lastname: Reichelderfer
mesh_terms: D000704:Analysis of Variance; D005069:Evaluation Studies as Topic; D015658:HIV Infections / Q000097:blood / Q000175:diagnosis*; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D012367:RNA, Viral / Q000032:analysis*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012661:Semen / Q000821:virology*; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic