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.
| Original language | English |
|---|---|
| Journal | Journal of Virological Methods |
| Volume | 169 |
| Issue number | 1 |
| Pages (from-to) | 70-78 |
| Number of pages | 9 |
| ISSN | 0166-0934 |
| DOIs | |
| Publication status | Published - Oct 2010 |
Keywords
- Animals
- Calicivirus, Feline
- Hepatitis A virus
- Humans
- Mytilus edulis
- Norovirus
- Ostreidae
- RNA, Viral
- Reverse Transcriptase Polymerase Chain Reaction
- Sensitivity and Specificity
- Time Factors
- Virology
