Abstract
Selecting the best quantitative PCR assay is essential to detect human norovirus genome effectively from clinical and environmental samples because no cell lines have been developed to propagate this virus. The real-time PCR methods for noroviruses GI (4 assays) and GII (3 assays) were evaluated using wastewater (n = 70) and norovirus-positive stool (n = 77) samples collected in Japan between 2012 and 2013. Standard quantitative PCR assays recommended by the U.S. Environmental Protection Agency, International Organization for Standardization, and Ministry of Health, Labour and Welfare, Japan, together with recently reported assays were included. Significant differences in positive rates and quantification cycles were observed by non-parametric analysis. The present study identifies the best assay for norovirus GI and GII to amplify norovirus genomes efficiently.
Highlights
Noroviruses (Family Caliciviridae, Genus Norovirus) are the leading etiologic agents causing viral gastroenteritis, with sporadic cases and outbreaks reported worldwide[1]
Most noroviruses used in the analysis were genotyped as GII.4 (59/77, 77%)
For GI assays in wastewater, the lowest positive rate was obtained with GI-B (p = 0.0002, p < 0.0001, and p = 0.0002 vs GI-A, GI-C, and GI-D, respectively)
Summary
Noroviruses (Family Caliciviridae, Genus Norovirus) are the leading etiologic agents causing viral gastroenteritis, with sporadic cases and outbreaks reported worldwide[1]. Because of its rapid molecular evolution, new genotypes and variants of the virus are reported frequently, and over 35 norovirus genotypes have been identified [2]. It is important to constantly ascertain the efficiency of the detection methods for the emerging genotypes. Despite great efforts by researchers [3], no cell line is currently available to propagate human norovirus. Molecular methods using PCR and quantitative real-time PCR (qPCR) are used widely to detect noroviruses in various fields including diagnostics, outbreak investigation, and environmental monitoring. Several qPCR methods have been developed and validated to detect noroviruses known to infect humans, namely genogroups I, II, and IV
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