title: Validation of an automated real-time PCR protocol for detection and quantitation of HIV and HCV genomes in semen.
abstract: The ability to detect and quantify HIV and HCV genomes is important for checking spermatozoid preparation protocols also known as "sperm washing". But no commercial assay is available. A method was developed for detecting HIV and HCV in semen fractions using the COBAS Ampliprep and COBAS Taqman instruments. It will detect fewer than 200 copies of HIV RNA per ml of semen plasma and fewer than 200 copies/3 x 10(6) semen cells. The sensitivity for HCV is similar at more than 200 IU/ml and below 200 IU/3 x 10(6) semen cells. No inhibitor of PCR amplification was detected. This automated protocol permits a convenient, standardized testing for HIV and HCV in semen. The performance is the same as that of the previous generation of automated assays but the cost and operating time are both reduced.
authors.affiliation: Laboratoire de Virologie, EA2046-IFR30, Hôpital Purpan, 31059 Toulouse, France. pasquier.c@chu-toulouse.fr
authors.forename: Christophe
authors.initials: C
authors.lastname: Pasquier
authors.forename: Corinne
authors.initials: C
authors.lastname: Souyris
authors.forename: Nathalie
authors.initials: N
authors.lastname: Moinard
authors.forename: Louis
authors.initials: L
authors.lastname: Bujan
authors.forename: Jacques
authors.initials: J
authors.lastname: Izopet
mesh_terms: D016679:Genome, Viral; D006678:HIV / Q000235:genetics / Q000302:isolation & purification*; D015658:HIV Infections / Q000821:virology; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000821:virology; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D012661:Semen / Q000821:virology*; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral