Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels

Katrine Uhrbrand, Mette Myrmel, Leena Maunula, Kirsti Vainio, Ramona Trebbien, Birgit Nørrung, Anna Charlotte Schultz

    33 Citationer (Scopus)

    Abstract

    Foodborne outbreaks caused by noroviruses (NoVs) and hepatitis A virus (HAV) are often linked to consumption of contaminated shellfish. The objective of this study was to identify an appropriate virus recovery method for real-time reverse transcriptase (RT)-PCR detection and subsequently to evaluate this method on shellfish bioaccumulated with virus in a collaborative study. Five methods were compared for recovery of NoV GII.7 and feline calicivirus from spiked digestive tissue of oysters and mussels. A method based on proteinase K digestion followed by NucliSENS miniMAG extraction was found to be the most efficient with a 50% limit of detection (LOD(50)) of 62 and 12 RT-PCR U/1.5 g digestive tissue for NoV GII.7 in oysters and mussels, respectively. Evaluation of the method in four laboratories found the percentage of sensitivity, based on low/high levels of virus bioaccumulated in oysters, to be 33/80 for NoV GI.3b, 13/92 for NoV GII.4 and 50/42 for HAV. A specificity of 100% was found for all three viruses in non-bioaccumulated oysters. As process control Mengovirus (vMC(0)) showed an average recovery of 1.8% from oysters and 1.2% from mussels. The study demonstrates that this recovery method can be useful for harmonized data generation and routine viral analyses of shellfish.
    OriginalsprogEngelsk
    TidsskriftJournal of Virological Methods
    Vol/bind169
    Udgave nummer1
    Sider (fra-til)70-78
    Antal sider9
    ISSN0166-0934
    DOI
    StatusUdgivet - okt. 2010

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