JCM Accepts, published online ahead of print on 21 October 2009

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J. Clin. Microbiol. doi:10.1128/JCM.00497-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

A Multi-center Comparison of Two Norovirus ORF2-based Genotyping Protocols

Kirsten Mattison^*, Elsie Grudeski, Brian Auk, Hugues Charest, Steven J. Drews, Angela Fritzinger, Nicole Gregoricus, Stephen Hayward, Alain Houde, Bonita E. Lee, Xiaoli L. Pang, Julie Wong, Tim F. Booth, and Jan Vinjé^*

Bureau of Microbial Hazards, Health Canada, Ottawa, ON; National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB; British Columbia Centre for Disease Control Laboratory Services, Vancouver, BC; Institut national de santé publique du Québec, Ste-Anne-de-Bellevue, QC; Ontario Agency for Health Protection and Promotion, Etobicoke, ON; Division of Consolidated Laboratory Services, Richmond, VA; Centers for Disease Control and Prevention, Atlanta, GA; Bureau of Biostatistics and Computer Applications, Health Canada, Ottawa, ON; Agriculture and Agri-Food Canada, Saint Hyacinthe, QC; Provincial Public Health Laboratory, Edmonton, AB

^* To whom correspondence should be addressed. Email: Kirsten_Mattison{at}hc-sc.gc.ca.

Norovirus outbreaks can be difficult to track due to the high frequency of environmental norovirus detection and the limited strain variation in some genotyping regions. However, rapid and accurate source identification can limit the spread of a foodborne outbreak and reduce the number of cases. Harmonization of genotyping assays is critical for enabling the rapid exchange of sequence data nationally and internationally. Several regions of the genome have been proposed for this purpose, but no consensus has been reached. In the present study, two standardized genotyping protocols (region C and region D) were evaluated by 9 laboratories in Canada and the United States using a coded panel of 96 fecal specimens representing 22 different norovirus genotypes. Overall, region C typing had a success rate of 78% compared to 52% for region D, however region D provides greater nucleotide sequence diversity for identifying new GII.4 variant strains. Significant differences in successful genotyping were observed between the 9 participating laboratories (10% to 100%) and between the different genotypes (6% to 100%). For several genogroup II strains, the reduced region D amplification correlated directly with mismatches between primer sequences and template. Based on overall performance, we recommended the region C protocol for routine genotyping of noroviruses while the region D protocol may be useful for identifying new GII.4 variants. Standardized genotyping will enable the rapid exchange of outbreak and sequence data through electronic norovirus surveillance networks.