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PubMed · 17184849

An RNA extraction protocol for shellfish-borne viruses.

David H Kingsley
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Abstract

The GPTT virus RNA extraction method, originally developed for extraction of human norovirus and hepatitis A virus RNAs from contaminated shellfish, was evaluated for extraction of RNA from Aichi virus strain A846/88 (AiV), coxsackievirus strains A9 (CAV9) and B5 (CBV5), murine norovirus (strain MNV-1), and the norovirus surrogate, feline calicivirus (FCV) strain KCD, for the purpose of RT-PCR detection within seeded oyster (Crassostrea virginica) extracts. The RT-PCR equivalent sensitivities observed within seeded oysters as compared to virus stocks were 0.68, 6.8, 26, 5.6, and 14.5 RT-PCR(50) units when assaying 10% of total RNA extracted from seeded oyster extracts for CAV9, CBV5, AiV, FCV, and MNV-1, respectively. For oysters exposed to virus-contaminated seawater, the detection equivalent sensitivities observed were 680, 68, 2600, 560, and 14.5 RT-PCR(50) for CAV9, CBV5, AiV and FCV, and MNV-1, respectively. These results indicate that the GPTT method can be used as a general viral RNA extraction method for multiple picornaviruses and caliciviruses that could potentially contaminate shellfish.

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AnimalsCaliciviridaeCatsEnterovirusEvaluation Studies as TopicKobuvirusNorovirusOstreidaeRNA, ViralReverse Transcriptase Polymerase Chain ReactionSensitivity and SpecificityShellfishEvaluation StudyJournal Article

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David H Kingsley. 2006-12-20. An RNA extraction protocol for shellfish-borne viruses.. https://doi.org/10.1016/j.jviromet.2006.11.027

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