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Journal of Clinical Microbiology, April 2003, p. 1423-1433, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1423-1433.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

International Collaborative Study To Compare Reverse Transcriptase PCR Assays for Detection and Genotyping of Noroviruses

Jan Vinjé,1,{dagger} Harry Vennema,1 Leena Maunula,2 Carl-Henrik von Bonsdorff,2 Marina Hoehne,3 Eckart Schreier,3 Alison Richards,4 Jon Green,4 David Brown,4 Suzanne S. Beard,5 Stephan S. Monroe,5 Erwin de Bruin,1 Lennart Svensson,6 and Marion P. G. Koopmans1*

Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, Bilthoven, The Netherlands,1 Haartman Institute, Department of Virology, Helsinki, Finland,2 Robert Koch-Institute, Berlin, Germany,3 Central Public Health Laboratory, London, United Kingdom,4 Viral Gastroenteritis Section, Centers for Disease Control and Prevention, Atlanta, Georgia,5 Swedish Institute for Infectious Disease Control, Solna, Sweden6

Received 6 May 2002/ Returned for modification 20 July 2002/ Accepted 29 December 2002

To allow more rapid and internationally standardized assessment of the spread of noroviruses (previously called Norwalk-like viruses [NLVs]) as important food-borne pathogens, harmonization of methods for their detection is needed. Diagnosis of NLVs in clinical diagnostic laboratories is usually performed by reverse transciptase PCR (RT-PCR) assays. In the present study, the performance of five different RT-PCR assays for the detection of NLVs was evaluated in an international collaborative study by five laboratories in five countries with a coded panel of 91 fecal specimens. The assays were tested for their sensitivity, detection limit, and ease of standardization. In total, NLVs could be detected by at least one RT-PCR assay in 69 (84%) of the samples that originally tested positive. Sensitivity ranged from 52 to 73% overall and from 54 to 100% and 58 to 85% for genogroup I and II viruses, respectively. In all, 64% of the false-negative results were obtained with a set of diluted stools (n = 20) that may have lost quality upon storage. Sensitivity was improved when these samples were excluded from analysis. No one single assay stood out as the best, although the p1 assay demonstrated the most satisfactory overall performance. To promote comparability of data, this assay will be recommended for newly starting groups in future collaborative studies.

* Corresponding author. Mailing address: Diagnostic Laboratory for Infectious Diseases and Perinatal Screening, National Institute of Public Health (RIVM), Bilthoven, The Netherlands. Phone: (31)30-2743945. Fax: (31)30-2744418. E-mail: marion.koopmans{at}rivm.nl.

{dagger} Present address: University of North Carolina, Chapel Hill, N.C.

Journal of Clinical Microbiology, April 2003, p. 1423-1433, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1423-1433.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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