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Journal of Clinical Microbiology, May 1999, p. 1409-1414, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Multicenter Quality Assessment of PCR Methods for Detection of Enteroviruses

Peter Muir,1,dagger ,* Albert Ras,2 Paul E. Klapper,3,dagger Graham M. Cleator,3,dagger Klaus Korn,4,dagger Christian Aepinus,5 Anders Fomsgaard,6 Pierre Palmer,7 Agneta Samuelsson,8 Antonio Tenorio,9 Benedikt Weissbrich,10,dagger and A. M. van Loon2,11,dagger

Department of Virology, Guy's, King's College & St Thomas' Hospitals' School of Medicine, London,1 and Division of Virology, Department of Pathological Sciences, Manchester Royal Infirmary, Manchester,3 United Kingdom; Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environmental (RIVM), Bilthoven,2 and Department of Virology, Utrecht Academic Hospital, Utrecht,11 The Netherlands; Institute for Clinical and Molecular Virology der Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen,4 Department of Molecular Pathology, University of Tübingen, Tübingen,5 and Institut für Virologie und Immunobiologie, Universität Würzburg, Würzburg,10 Germany; Department of Virology, Statens Seruminstitut, Copenhagen, Denmark6; Service de Bacteriologie, Virologie et Hygiene, Hôpital Saint Vincent de Paul, Paris, France7; Clinical Virology F68, Huddinge University Hospital, Huddinge, Sweden8; and C.N.M., Instituto de Salud Carlos III, Madrid, Spain9

Received 17 September 1998/Returned for modification 7 December 1998/Accepted 2 February 1999

We conducted a multicenter evaluation of commercial and in-house PCR methods for the detection of enteroviruses. Three coded panels of test and control RNA samples, artificial clinical specimens, and representative enterovirus serotypes were used to assess amplification methods, RNA extraction methods, and reactivities with different enterovirus serotypes. Despite several differences between PCR methods, there was good agreement, although some variation in sensitivity was observed. Most PCR methods were able to detect enterovirus RNA derived from 0.01 50% tissue culture infective dose (TCID50) and were able to detect at least 1 TCID50 of enterovirus in cerebrospinal fluid, stool, or throat swab specimens. Most were also able to detect a wide range of enterovirus serotypes, although serotypic identification was not possible. Some laboratories experienced false-positive results due to PCR contamination, which appeared to result mainly from cross-contamination of specimens during RNA extraction. Provided that this problem is overcome, these PCR methods will prove to be a sensitive and rapid alternative to cell culture for the diagnosis of enterovirus infection.

* Corresponding author. Mailing address: Department of Virology, Guy's, King's College & St Thomas' Hospitals' School of Medicine, St. Thomas' Campus, Lambeth Palace Rd., London SE1 7EH, United Kingdom. Phone: (44) 171 922 8167. Fax: (44) 171 922 8387. E-mail: p.muir{at}umds.ac.uk.

dagger Member of the European Union Concerted Action on Virus Meningitis and Encephalitis. Other members of the group are listed in the Appendix.

Journal of Clinical Microbiology, May 1999, p. 1409-1414, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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