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Journal of Clinical Microbiology, June 1998, p. 1741-1745, Vol. 36, No. 6
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Evaluation of a Commercially Available Reverse Transcription-PCR Assay for Diagnosis of Enteroviral Infection in Archival and Prospectively Collected Cerebrospinal Fluid Specimens

Francisco Pozo,1,* Inmaculada Casas,1 Antonio Tenorio,1 Gloria Trallero,2 and Jose M. Echevarria1,dagger

Diagnostic Microbiology Service1 and Virology Service,2 Centro Nacional de Microbiología, Majadahonda, Madrid, Spain

Received 10 November 1997/Returned for modification 15 January 1998/Accepted 18 March 1998

A commercially available reverse transcription (RT)-PCR method (AMPLICOR EV; Roche Diagnostic Systems, Inc., Branchburg, N.J.) was evaluated for detection of enteroviruses in cerebrospinal fluid from patients with neurological disease. This assay was compared with virus isolation in cell culture and an in-house RT-PCR method designed with a nonoverlapping region of the enteroviral genome. A panel of 200 cerebrospinal fluid specimens prospectively collected from patients with a wide variety of neurological symptoms, including 50 patients involved in three different outbreaks of acute aseptic meningitis, was assayed. A second panel of 97 archived cerebrospinal fluid specimens, stored for 2 to 5 years, from patients with aseptic meningitis associated with several enterovirus outbreaks was also studied. From the first panel, enteroviruses were detected in 13 of 50 specimens by cell culture (26%), in 43 of 50 specimens by AMPLICOR EV (86%), and in 46 of 50 specimens by the in-house assay (92%) from patients with aseptic meningitis associated with outbreak and 1 of 29, 3 of 29, and 4 of 29 specimens, respectively, from sporadic cases of aseptic meningitis. The remaining 121 cerebrospinal fluid specimens from patients with other neurological syndromes were negative by all tests. From the second panel, enteroviral RNA was detected by the AMPLICOR test (31 of 97 specimens, 32%) and the in-house assay (39 of 97 specimens, 40%). According to our results, patients with aseptic meningitis should be analyzed for enteroviral infection in cerebrospinal fluid by RT-PCR methods, and the AMPLICOR EV test is a suitable tool for performing such studies. Archival cerebrospinal fluid specimens are less suitable for evaluation of the performance of RT-PCR methods designed for enterovirus detection.

* Corresponding author. Mailing address: Servicio de Microbiología Diagnóstica, Centro Nacional de Microbiología, Ctra. de Pozuelo Km 2, 28220 Majadahonda, Madrid, Spain. Phone: 34 1 509 79 01. Fax: 34 1 509 79 66. E-mail: jmecheva{at}isciii.es.

dagger Investigator representing the European Union Concerted Action on Virus Meningitis and Encephalitis group. Other group members are G. M. Cleator, Department of Pathological Sciences, University of Manchester, Manchester, United Kingdom; Maria Ciardi, Universita di Roma `La Sapienza,' Rome, Italy; Paola Cinque, Ospedale San Raffaele, Milan, Italy; José Manuel Echevarria, Instituto de Salud Carlos III, Madrid, Spain; Marianne Forsgren, Huddinge Hospital, Stockholm, Sweden; Giuseppe Gerna, IRCCS Policlinico San Matteo, Pavia, Italy; Monica Grandien, Swedish Institute for Infectious Disease Control, Stockholm, Sweden; Frauke Harms, Universität Würzburg, Würzburg, Germany; Tapani Hovi, National Public Health Institute, Helsinki, Finland; Paul Klapper, Manchester Royal Infirmary, Manchester, United Kingdom; Marjaleena Koskiniemi, University of Helsinki, Helsinki, Finland; Pierre Lebon, Hôpital Saint Vincent de Paul, Paris, France; Annika Linde, Swedish Institute for Infectious Disease Control, Stockholm, Sweden; Anton van Loon, Academic Hospital Utrecht, Utrecht, The Netherlands; Volker ter Meulen, Universität Würzburg, Würzburg, Germany; Philippe Monteyne, Université Catholique de Louvain, Brussels, Belgium; Peter Muir, UMDS Guys & St. Thomas' Hospitals, London, United Kingdom; Elisabeth Puchhammer-Stöckl, University of Vienna, Vienna, Austria; Floré Rozenberg, Hôpital Saint Vincent de Paul, Paris, France; Christian Sindic, Université Catholique de Louvain, Brussels, Belgium; Clive Taylor, Newcastle General Hospital, Newcastle-upon-Tyne, United Kingdom; Bent Vestergaard, Statens Seruminstitut, Copenhagen, Denmark; Thomas Weber, Marienkrankenhaus Hamburg, Hamburg, Germany; and Benedikt Weissbrich, Universität Würzburg, Würzburg, Germany.

Journal of Clinical Microbiology, June 1998, p. 1741-1745, Vol. 36, No. 6
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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