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Estimation of Human Norovirus Infectivity from Environmental Water Samples by In Situ Capture RT-qPCR Method

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Abstract

Human noroviruses (HuNoVs) are highly infectious viruses for which water is an important medium of transmission. In this study, we explored a new in situ capture RT-qPCR (ISC-RT-qPCR) methodology to estimate the infectivity of HuNoV in environmental water samples. This assay was based on capturing encapsidated HuNoV by viral receptors, followed by in situ amplification of the captured viral genomes by RT-qPCR. We demonstrated that the ISC-RT-qPCR did not capture and enable signal amplification of heat-denatured Tulane Virus (TV) and HuNoVs. We further demonstrated that the sensitivity of ISC-RT-qPCR was equal or better than that of conventional RT-qPCR procedures for the detection of HuNoV GI and GII. We then utilized the ISC-RT-qPCR to detect HuNoV in environmental water samples for comparison against that from a conventional RT-qPCR procedure. TV was used as a process control virus. While complete inhibition of TV genomic signal was observed in 27% of samples tested by RT-qPCR, no inhibition of TV genomic signal was observed by ISC-RT-qPCR. From 72 samples tested positive for HuNoV GI signal by RT-qPCR, only 20 (27.8%) of these samples tested positive by ISC-RT-qPCR, suggesting that 72.2% of RT-qPCR-positive samples were unlikely to be infectious. From 16 samples tested positive for HuNoV GII signal by RT-qPCR, only one of these samples tested positive by ISC-RT-qPCR. Five samples that had initially tested negative for HuNoV GII signal by RT-qPCR, was tested as positive by ISC-RT-qPCR. Overall, ISC-RT-qPCR method provided an alternative assay to estimate infectivity of HuNoV in environmental samples.

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Acknowledgements

This work was supported by USDA Agricultural Research Service CRIS project 5325-42000-050-00D and Grant 31301475 from National Natural Science Foundation of China. USDA is an equal opportunity provider and employer.

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Authors and Affiliations

  1. Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, 800 Buchanan St., Albany, CA, 94710-1105, USA

    Peng Tian, David Yang & Lei Shan

  2. Department of Bioengineering, Shanghai Institute of Technology, Shanghai, 100 Haiquan Road, Fengxian District, Shanghai, 201418, People’s Republic of China

    Qianqian Li

  3. MOST-USDA Joint Research Center for Food Safety & Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People’s Republic of China

    Danlei Liu & Dapeng Wang

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  1. Peng Tian
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Correspondence to Peng Tian.

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Tian, P., Yang, D., Shan, L. et al. Estimation of Human Norovirus Infectivity from Environmental Water Samples by In Situ Capture RT-qPCR Method. Food Environ Virol 10, 29–38 (2018). https://doi.org/10.1007/s12560-017-9317-1

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