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Development and validation of a microarray for the confirmation and typing of norovirus RT-PCR products |
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| Authors: | Mattison Kirsten Corneau Nathalie Berg Ingvild Bosch Albert Duizer Erwin Gutiérrez-Aguirre Ion L'homme Yvan Lucero Yalda Luo Zhiyao Martyres Alexander Myrmel Mette O'Ryan Miguel Pagotto Franco Sano Daisuke Svraka Sanela Urzua Ulises Bidawid Sabah |
| Affiliation: | a Bureau of Microbial Hazards, Health Canada, Ottawa, Canada b Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Canada c Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, Oslo, Norway d Department of Microbiology, University of Barcelona, Barcelona, Spain e Department of Virology, National Institute for Public Health and the Environment, Bilthoven, Netherlands f Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia g St-Hyacinthe Laboratory, Canadian Food Inspection Agency, St-Hyacinthe, Canada h Faculty of Medicine, University of Chile, Santiago, Chile |
| Abstract: | Noroviruses are implicated in many worldwide institutional, food and waterborne outbreaks each year. Genetic typing of isolates is valuable for monitoring outbreak spread as well as variation in circulating strains. Microarrays have the potential to provide rapid genotype information for norovirus samples. The NoroChip v3.0 provides an oligonucleotide hybridization platform to screen for over 600 potential interactions in each experiment. The NoroChip v3.0 was developed at Health Canada and validated in seven international partner laboratories. Each laboratory validated the NoroChip v3.0 using norovirus amplicons routinely characterized in their testing protocols. Fragments from the capsid region (region C) and a 2.4 kb amplicon spanning polymerase and capsid sequences (region AD) were validated in six of the partner laboratories and provided correct genogroup typing information (GI or GII) when hybridized to the NoroChip v3.0. Results indicate that the current limiting factor for implementing the NoroChip v3.0 as a strain typing tool is the difficulty obtaining a long, specific amplicon from all circulating norovirus strains. Data obtained with the longer region AD amplicon provided the best discrimination between norovirus strains. |
| Keywords: | Norovirus Microarray Genotyping |
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