title: Multicenter quality control for the detection of hepatitis C virus RNA in seminal plasma specimens.
abstract: The discrepant results available in the literature about the presence of hepatitis C virus (HCV) RNA in seminal plasma of men chronically infected by this agent are related, at least in part, to the molecular techniques used and particularly to the wide range of protocols dedicated to RNA extraction. In order to evaluate these protocols and to standardize the method of detection of HCV RNA in this fluid, a panel of coded specimens was tested blindly in 12 French laboratories; it included 14 seminal plasma specimens and four water controls spiked with HCV RNA ranging from 10 to 20000 IU/ml and two HCV-negative seminal plasma specimens. The extraction step was performed according to methods using either silica beads (NucliSens [Organon Teknika S.A., Fresnes, France]; RNA viral kit [Qiagen, Courtaboeuf, France]) or guanidinium thiocyanate (Amplicor HCV assay; Roche Diagnostics, Meylan, France), preceded or not by a centrifugation of the seminal plasma. For the amplification step, all the laboratories performed the same reverse transcription-PCR technique (Amplicor HCV Cobas assay). The percentage of correct results ranged from 53.3 to 100, the poorest results being obtained when no centrifugation step preceded the Amplicor extraction protocol. The rate of correct results was significantly higher in laboratories using a preliminary centrifugation of the specimen (P = 0.034 by chi-square test). By contrast, the overall number of correct results was not correlated to the initial volume of sample used for the test. These results allowed us to validate standardized techniques adapted to the performance of this test on a routine basis, especially in men infected with HCV and involved in programs of medically assisted reproduction.
authors.affiliation: Laboratoire de Bactériologie-Virologie, Unité de Pharmacologie Clinique, Faculté de Médecine, University of Saint-Etienne, Saint-Etienne, France. Thomas.Bourlet@univ-st-etienne.fr
authors.forename: Thomas
authors.initials: T
authors.lastname: Bourlet
authors.forename: Rachel
authors.initials: R
authors.lastname: Levy
authors.forename: Silvy
authors.initials: S
authors.lastname: Laporte
authors.forename: Stéphane
authors.initials: S
authors.lastname: Blachier
authors.forename: Laurence
authors.initials: L
authors.lastname: Bocket
authors.forename: Guy
authors.initials: G
authors.lastname: Cassuto
authors.forename: Lionel
authors.initials: L
authors.lastname: Chollet
authors.forename: Marianne
authors.initials: M
authors.lastname: Leruez-Ville
authors.forename: Anne
authors.initials: A
authors.lastname: Maertens
authors.forename: Fabienne
authors.initials: F
authors.lastname: Mousnier
authors.forename: Christophe
authors.initials: C
authors.lastname: Pasquier
authors.forename: Christopher
authors.initials: C
authors.lastname: Payan
authors.forename: Bertrand
authors.initials: B
authors.lastname: Pellegrin
authors.forename: Evelyne
authors.initials: E
authors.lastname: Schvoerer
authors.forename: Patricia
authors.initials: P
authors.lastname: Zavadzki
authors.forename: Jacques
authors.initials: J
authors.lastname: Chouteau
authors.forename: Gilles
authors.initials: G
authors.lastname: Duverlie
authors.forename: Jacques
authors.initials: J
authors.lastname: Izopet
authors.forename: Françoise
authors.initials: F
authors.lastname: Lunel-Fabiani
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Pawlotsky
authors.forename: Nerina
authors.initials: N
authors.lastname: Profizi
authors.forename: Christine
authors.initials: C
authors.lastname: Rouzioux
authors.forename: Françoise
authors.initials: F
authors.lastname: Stoll-Keller
authors.forename: Vincent
authors.initials: V
authors.lastname: Thibault
authors.forename: Pierre
authors.initials: P
authors.lastname: Wattré
authors.forename: Bruno
authors.initials: B
authors.lastname: Pozzetto
mesh_terms: D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D011786:Quality Control; D012367:RNA, Viral / Q000032:analysis*; D012661:Semen / Q000821:virology*
chemical_list: D012367:RNA, Viral