Multicenter quality control for the detection of hepatitis C virus RNA in seminal plasma specimens.
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.
Bourlet
Thomas
T
Laboratoire de Bactériologie-Virologie, Unité de Pharmacologie Clinique, Faculté de Médecine, University of Saint-Etienne, Saint-Etienne, France. Thomas.Bourlet@univ-st-etienne.fr
Levy
Rachel
R
Laporte
Silvy
S
Blachier
Stéphane
S
Bocket
Laurence
L
Cassuto
Guy
G
Chollet
Lionel
L
Leruez-Ville
Marianne
M
Maertens
Anne
A
Mousnier
Fabienne
F
Pasquier
Christophe
C
Payan
Christopher
C
Pellegrin
Bertrand
B
Schvoerer
Evelyne
E
Zavadzki
Patricia
P
Chouteau
Jacques
J
Duverlie
Gilles
G
Izopet
Jacques
J
Lunel-Fabiani
Françoise
F
Pawlotsky
Jean-Michel
JM
Profizi
Nerina
N
Rouzioux
Christine
C
Stoll-Keller
Françoise
F
Thibault
Vincent
V
Wattré
Pierre
P
Pozzetto
Bruno
B
D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D011786:Quality Control; D012367:RNA, Viral / Q000032:analysis*; D012661:Semen / Q000821:virology*
D012367:RNA, Viral