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Effect of the Shellfish Proteinase K Digestion Method on Norovirus Capsid Integrity

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Abstract

Norovirus outbreaks are associated with the consumption of contaminated shellfish, and so efficient methods to recover and detect infectious norovirus in shellfish are important. The Proteinase K digestion method used to recover norovirus from shellfish, as described in the ISO 15216, would be a good candidate but its impact on the virus capsid integrity and thus infectivity was never examined. The aim of this study was to assess the impact of the Proteinase K digestion method, and of the heat treatment component of the method alone, on norovirus (genogroups I and II) and MS2 bacteriophage capsid integrity. A slightly modified version of the ISO method was used. RT-qPCR was used for virus detection following digestion of accessible viral RNA using RNases. MS2 phage infectivity was measured using a plaque assay. The effect of shellfish digestive glands (DG) on recovery was evaluated. In the presence of shellfish DG, a reduction in MS2 phage infectivity of about 1 log10 was observed after the Proteinase K digestion method and after heat treatment component alone. For norovirus GII and MS2 phage, there was no significant loss of genome following the Proteinase K digestion method but there was a significant 0.24 log10 loss of norovirus GI. In the absence of shellfish DG, the reduction in MS2 phage infectivity was about 2 log10, with the addition of RNases resulting in a significant loss of genome for all tested viruses following complete Proteinase K digestion method and the heat treatment alone. While some protective effect from the shellfish DG on viruses was observed, the impact on capsid integrity and infectivity suggests that this method, while suitable for norovirus genome detection, may not completely preserve virus infectivity.

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Acknowledgements

This work was funded by the Ministry of Business, Innovation and Employment (MBIE) (Safe New Zealand Seafood. CAWX1317 Project 15330). The authors would like to thank Dr Gail Greening and Dr Pradip Gyawali for their critical review of the manuscript. Thanks also to Jim Dollimore for kindly providing Pacific oysters.

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  1. Enteric, Environmental and Food Virology Laboratory, Institute of Environmental Science and Research Ltd (ESR), Kenepuru Science Centre, PO BOX 50-348, Porirua, 5240, New Zealand

    Jérémie Langlet, Laetitia Kaas, Dawn Croucher & Joanne Hewitt

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  1. Jérémie Langlet
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  2. Laetitia Kaas
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Correspondence to Jérémie Langlet.

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Langlet, J., Kaas, L., Croucher, D. et al. Effect of the Shellfish Proteinase K Digestion Method on Norovirus Capsid Integrity. Food Environ Virol 10, 151–158 (2018). https://doi.org/10.1007/s12560-018-9336-6

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  • DOI: https://doi.org/10.1007/s12560-018-9336-6

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