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Water 2018, 10(7), 841; https://doi.org/10.3390/w10070841

Detection of Infectious Noroviruses from Wastewater and Seawater Using PEMAXTM Treatment Combined with RT-qPCR

Institute of Environmental Science and Research Ltd. (ESR), Porirua 5240, New Zealand
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Received: 19 May 2018 / Revised: 14 June 2018 / Accepted: 22 June 2018 / Published: 25 June 2018
(This article belongs to the Special Issue Health Risks of Alternative Water Sources)
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Abstract

Rapid detection of infectious noroviruses from environmental samples is essential to minimize the risk of norovirus outbreaks associated with environmental transmission. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) methods are rapid and sensitive, but cannot differentiate between infectious and non-infectious noroviruses. In this study, a PEMAXTM treatment followed by RT-qPCR (PEMAXTM-RT-qPCR) method was developed for murine norovirus and norovirus GI/GII, and evaluated for the selective detection of infectious viruses following heat inactivation. The norovirus PEMAXTM-RT-qPCR method was then evaluated for the selective detection of infectious viruses from environmental samples. Following heat-treatment (90 °C for 3 min), the murine norovirus PEMAXTM-RT-qPCR showed at least a 2.04 log10 reduction in detectable virus, compared to a 0.43 log10 reduction for RT-qPCR alone. Under the same conditions, the norovirus PEMAXTM-RT-qPCR showed a 0.34 to 0.98 log10 (GI.3) and 0.63 to 2.06 log10 (GII.4) reduction in detectable viruses, compared to 0.05 to 0.18 log10 (GI.3) and 0.06 to 0.25 log10 (GII.4) for RT-qPCR alone. Evaluation of the norovirus PEMAXTM-RT-qPCR on norovirus-contaminated influent and effluent wastewater, and seawater indicated a high proportion of non-infectious norovirus GI and GII (i.e., 56 to 100% in seawater, 32 to 76% in effluent, and 11 to 79% in influent) was present in samples. While potentially overestimating the amount of infectious noroviruses, this approach has potential to provide better information on viral infectivity than RT-qPCR alone. View Full-Text
Keywords: norovirus; PEMAXTM-RT-qPCR; infectivity assessment; environmental water; health risks norovirus; PEMAXTM-RT-qPCR; infectivity assessment; environmental water; health risks
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Gyawali, P.; Hewitt, J. Detection of Infectious Noroviruses from Wastewater and Seawater Using PEMAXTM Treatment Combined with RT-qPCR. Water 2018, 10, 841.

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