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Journal article

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

By Uhrbrand, Katrine1,2; Myrmel, Mette3; Maunula, Leena4; Vainio, Kirsti5; Trebbien, Ramona6,7,8; Nørrung, Birgit2; Schultz, Anna Charlotte1,2

From

Division of Microbiology and Risk Assessment, National Food Institute, Technical University of Denmark1

National Food Institute, Technical University of Denmark2

Norwegian School of Veterinary Science3

University of Helsinki4

Norwegian Institute of Public Health5

Virology, Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark6

Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark7

National Veterinary Institute, Technical University of Denmark8

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 (LOD50) 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 (vMC0) 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.

Language: English
Year: 2010
Pages: 70-78
ISSN: 18790984 and 01660934
Types: Journal article
DOI: 10.1016/j.jviromet.2010.06.019
ORCIDs: Uhrbrand, Katrine and Schultz, Anna Charlotte
Keywords

Hepatitis A virus Mussel Norovirus Oyster Real-time RT-PCR detection SDG 3 - Good Health and Well-being Virus recovery

Other keywords

Animals Calicivirus, Feline Humans Mytilus edulis Ostreidae RNA, Viral Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Time Factors Virology

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