Detection and Quantification of Noroviruses in Shellfish

Type Article
Date 2009-02
Language English
Author(s) Le Guyader Soizick1, Parnaudeau Sylvain1, Schaeffer Julien1, Bosch Albert2, Loisy Fabienne3, Pommepuy Monique1, Atmar Robert L.4
Affiliation(s) 1 : IFREMER, Microbiol Lab, F-44311 Nantes 03, France.
2 : Univ Barcelona, Dept Microbiol, Enter Virus Lab, Barcelona, Spain.
3 : CEERAM SAS, La Chapelle Sur Erdre, France.
4 : Baylor Coll Med, Dept Mol Virol & Microbiol, Houston, TX 77030 USA.
Source Applied and environmental microbiology (0099-2240) (American society for microbiology), 2009-02 , Vol. 75 , N. 3 , P. 618-624
DOI 10.1128/AEM.01507-08
WOS© Times Cited 149
Abstract Noroviruses (NoVs) are the most common viral agents of acute gastroenteritis in humans, and high concentrations of NoVs are discharged into the environment. As these viruses are very resistant to inactivation, the sanitary consequences are contamination of food, including molluscan shellfish. There are four major problems with NoV detection in shellfish samples: low levels of virus contamination, the difficulty of efficient virus extraction, the presence of interfering substances that inhibit molecular detection, and NoV genetic variability. The aims of this study were to adapt a kit for use with a method previously shown to be efficient for detection of NoV in shellfish and to use a one step real-time reverse transcription-PCR method with addition of an external viral control. Comparisons of the two methods using bioaccumulated oysters showed that the methods reproducibly detected similar levels of virus in oyster samples. Validation studies using naturally contaminated samples also showed that there was a good correlation between the results of the two methods, and the variability was more attributable to the level of sample contamination. Magnetic silica very efficiently eliminated inhibitors, and use of extraction and amplification controls increased quality assurance. These controls increased the confidence in estimates of NoV concentrations in shellfish samples and strongly supported the conclusion that the results of the method described here reflected the levels of virus contamination in oysters. This approach is important for food safety and is under evaluationfor European regulation.
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Le Guyader Soizick, Parnaudeau Sylvain, Schaeffer Julien, Bosch Albert, Loisy Fabienne, Pommepuy Monique, Atmar Robert L. (2009). Detection and Quantification of Noroviruses in Shellfish. Applied and environmental microbiology, 75(3), 618-624. Publisher's official version : , Open Access version :