Comparative Evaluation of Real-Time PCR Methods for Human Noroviruses in Wastewater and Human Stool

PLoS One. 2016 Aug 15;11(8):e0160825. doi: 10.1371/journal.pone.0160825. eCollection 2016.

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.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Child
  • Feces / virology*
  • Genotype
  • Humans
  • Norovirus / genetics*
  • Norovirus / isolation & purification*
  • Real-Time Polymerase Chain Reaction / methods*
  • Statistics, Nonparametric
  • Wastewater / virology*

Substances

  • Waste Water

Grants and funding

Support was provided by Japan Science and Technology Agency; Core Research for Evolutional Science and Technology [http://www.jst.go.jp/kisoken/crest/en/] to TO; and Japan Society for the Promotion of Science; Grant-in-Aid for Scientific Research (15H04065) [https://www.jsps.go.jp/english/e-grants/] to YM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.