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Simplified Procedure for Detection of Poliovirus and Norovirus in Oysters  

Ha, Sook-Hee (Department of Biotechnology, Dongguk University)
Woo, Gun-Jo (Center for Food Safety Evaluation, Korea Food and Drug Administration)
Kwak, Hyo-Sun (Center for Food Safety Evaluation, Korea Food and Drug Administration)
Hwang, In-Gyun (Center for Food Safety Evaluation, Korea Food and Drug Administration)
Choi, Weon-Sang (Department of Biotechnology, Dongguk University)
Publication Information
Korean Journal of Food Science and Technology / v.37, no.6, 2005 , pp. 1018-1023 More about this Journal
Abstract
Simplified procedure was developed for concentrating and detecting poliovirus and norovirus in oysters. Viruses were seeded into oyster tissue homogenates and concentrated through polyethylene glycol (PEG) precipitation, chloroform or Freon extraction, with additional PEG precipitation. Amount of viruses was evaluated using poliovirus plaque assay. Virus recovery during concentration procedure was approximately 16.4-26.0%. For defection, viral RNAs in oysters were examined using one-step RT-PCR after extraction with Trizol. Dilution or capturing of viral RNA using silica gel membrane allowed viruses to be detected by RT-PCR, whereas viruses could not be removed using $QIAshredder^{TM}$ Homogenizer, which is effective in removing RT-PCR inhibitors in lettuce and hamburgers. Freon extraction, generally used to concentrate viruses found in food, could be substituted with chloroform extraction using this procedure; no difference could be observed between detection limits of whole oyster extracts and digestive organ extracts indicating that RT-PCR inhibitors were distributed evenly throughout whole tissues. Nested PCR greatly improved efficiency of this procedure. Overall, this procedure could remove sufficient amount of inhibitors to allow detection of norovirus in oysters.
Keywords
poliovirus; norovirus; oyster; RT-PCR;
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