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KMID : 0613820020120010049
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Journal of Life Science
2002 Volume.12 No. 1 p.49 ~ p.54
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Development of a simple and sensitive method to detect enteric viruses from oysters
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Jung Eun-Young
Cheon Doo-Sung
Jee Young-Mee
Yoon Jae-Deuk
Cho Hae-Wol
Jang Kyung-Lib
Je Hee Bok
Jun Hong-Ki
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Abstract
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Development of a rapid method possessing the requisite sensitivity and specificity for virus monitoring is necessary for protection of the shellfish-consuming public. Oysters tissue usually contains virus particles in relatively small concentrations along with various other substances that can interfere with detection steps. Therefore, the critical point concerning the detection of viruses in shellfish tissues resides in the processing of samples. The current study demonstrated the possibility of purifying small amounts of virus particles at the interface of a 10/50% sucrose
gradient after a single round of sucrose gradient ultracentrifugation. We could detect HAV and poliovirus simultaneously from oyster tissues by using two different sets of primer. Furthermore, the method showed a high level of virus recovery rate (>95%) as determined by plaque assays of the final samples. Taken the advantages of the simple and sensitive methods, it was possible to detect 2 pfu of HAV in 5 g of oyster digestive tissues within 24h.
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KEYWORD
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enteric viruses, hepatitis A virus, oyster, RT-PCR, gradient-ultracentrifugation
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