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A duplex real-time RT-PCR assay for the simultaneous genogroup-specific detection of noroviruses in both clinical and environmental specimens

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Abstract

Norovirus (NoV) is the major etiological agent causing foodborne and waterborne outbreaks worldwide. We developed a novel duplex real-time quantitative RT-PCR assay designed for the simultaneous detection of and discrimination between NoV genogroups GI and GII, by targeting the short junction region between ORF1 and ORF2, with sensitivity and efficiency comparable to those of each simplex RT-PCR assay. This new duplex assay was evaluated against clinical stool (n = 82) and environmental (groundwater or surface water, n = 60) specimens from South Korea, and the results were compared with those of conventional RT-PCR (cRT-PCR) assays. The duplex assay detected more positive samples than did the cRT-PCR for both clinical (74 vs. 71) and, more strikingly, environmental (24 vs. 10) specimens. No cross-reactivity against specimens containing other enteric viruses such as rotavirus, adenovirus, and poliovirus were observed. These results suggest that this newly developed duplex real-time RT-PCR assay can be used for the sensitive and simultaneous genogroup-specific detection of NoV in both clinical and environmental specimens.

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Acknowledgment

This research was supported by the Agriculture Research Center program of the Ministry of Food, Agriculture, Forestry, and Fisheries, and National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (2011-0000177).

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Authors and Affiliations

  1. Department of Environmental Health and Institute of Health and Environment, School of Public Health, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 151-742, South Korea

    YoungBin Park, You-Hee Cho & GwangPyo Ko

  2. Center for Food Safety and Toxicology, Seoul National University, Seoul, Korea

    GwangPyo Ko

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  1. YoungBin Park
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  2. You-Hee Cho
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Correspondence to GwangPyo Ko.

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Park, Y., Cho, YH. & Ko, G. A duplex real-time RT-PCR assay for the simultaneous genogroup-specific detection of noroviruses in both clinical and environmental specimens. Virus Genes 43, 192–200 (2011). https://doi.org/10.1007/s11262-011-0626-4

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  • DOI: https://doi.org/10.1007/s11262-011-0626-4

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