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Journal of Clinical Microbiology, February 2003, p. 789-793, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.789-793.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Multicenter Quality Control for the Detection of Hepatitis C Virus RNA in Seminal Plasma Specimens

Thomas Bourlet,^1* Rachel Levy,² Silvy Laporte,³ Stéphane Blachier,⁴ Laurence Bocket,⁵ Guy Cassuto,⁶ Lionel Chollet,⁷ Marianne Leruez-Ville,⁸ Anne Maertens,¹ Fabienne Mousnier,⁹ Christophe Pasquier,¹⁰ Christopher Payan,¹¹ Bertrand Pellegrin,¹² Evelyne Schvoerer,¹³ Patricia Zavadzki,¹⁴ Jacques Chouteau,⁴ Gilles Duverlie,¹⁴ Jacques Izopet,¹⁰ Françoise Lunel-Fabiani,¹¹ Jean-Michel Pawlotsky,¹² Nerina Profizi,⁷ Christine Rouzioux,⁸ Françoise Stoll-Keller,¹³ Vincent Thibault,⁹ Pierre Wattré,⁵ and Bruno Pozzetto¹

Laboratoire de Bactériologie-Virologie,¹ Laboratoire de Biologie de la Reproduction,² Unité de Pharmacologie Clinique, Faculté de Médecine, University of Saint-Etienne, Saint-Etienne,³ Laboratoire Clinilab, Saint-Ismier,⁴ Laboratoire de Virologie, CHU de Lille, Lille,⁵ LABM Drouot,⁶ Laboratoire de Microbiologie, Hôpital Necker,⁸ Laboratoire de Virologie-CERVI, Hôpital de La Pitié Salpétrière, Paris,⁹ Laboratoire de Virologie, Hôpital Inter-communal, Toulon-La Seyne sur Mer,⁷ Laboratoire de Virologie, Hôpital Purpan, Toulouse,¹⁰ Laboratoire de Virologie, CHU d'Angers, Angers,¹¹ Laboratoire de Bactériologie-Virologie, Hôpital H. Mondor, Créteil,¹² Laboratoire de Bactériologie-Virologie, Faculté de Médecine, University of Strasbourg, Strasbourg,¹³ Laboratoire de Virologie, CHU d'Amiens, Amiens, France,¹⁴

Received 9 May 2002/ Returned for modification 5 August 2002/ Accepted 20 November 2002

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 20,000 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.

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* Corresponding author. Mailing address: Department of Bacteriology-Virology GIMAP, Faculty of Medicine, University of Saint-Etienne, 15 rue A. Paré, 42023 Saint-Etienne Cedex, France. Phone: (33) 4 77 82 83 15. Fax: (33) 4 77 82 84 60. E-mail: Thomas.Bourlet{at}univ-st-etienne.fr.

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Journal of Clinical Microbiology, February 2003, p. 789-793, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.789-793.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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