Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels.
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.
Uhrbrand
Katrine
K
Division of Microbiology and Risk Assessment, National Food Institute, Technical University of Denmark, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark.
Myrmel
Mette
M
Maunula
Leena
L
Vainio
Kirsti
K
Trebbien
Ramona
R
Nørrung
Birgit
B
Schultz
Anna Charlotte
AC
D000818:Animals; D017927:Calicivirus, Feline / Q000302:isolation & purification; D030041:Hepatitis A virus / Q000302:isolation & purification*; D006801:Humans; D049880:Mytilus edulis / Q000821:virology*; D029322:Norovirus / Q000302:isolation & purification*; D010125:Ostreidae / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013997:Time Factors; D014773:Virology / Q000379:methods*
D012367:RNA, Viral