Simplified Procedure for Detection of Poliovirus and Norovirus in Oysters

굴로부터 오염된 폴리오바이러스 및 노로바이러스의 검출

  • 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)
  • 하숙희 (동국대학교 생명공학과) ;
  • 우건조 (식품의약품안전청 식품안전평가부) ;
  • 곽효선 (식품의약품안전청 식품안전평가부) ;
  • 황인균 (식품의약품안전청 식품안전평가부) ;
  • 최원상 (동국대학교 생명공학과)
  • Published : 2005.12.31

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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