Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Development of a simple and sensitive method to detect enteric viruses from oysters

굴로부터 장바이러스를 검출하기 위한 간단하고 민감한 방법의 개발

  • Chung, Eun-Young (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Je, Hee-Bok (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Jun, Hong-Ki (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Yoon, Jae-Deuk (Laboratory of Enteroviruses, Department of Virology, National Institute of Health) ;
  • Jee, Young-Mee (Laboratory of Enteroviruses, Department of Virology, National Institute of Health) ;
  • Cheon, Doo-Sung (Laboratory of Enteroviruses, Department of Virology, National Institute of Health) ;
  • Cho, Hae-Wol (Laboratory of Enteroviruses, Department of Virology, National Institute of Health) ;
  • Jang, Kyung-Lib (Department of Microbiology, College of Natural Sciences, Pusan National University)
  • 정은영 (부산대학교 미생물학과) ;
  • 제회복 (부산대학교 미생물학과) ;
  • 전홍기 (부산대학교 미생물학과) ;
  • 윤재득 (국립보건원 바이러스과) ;
  • 지영미 (국립보건원 바이러스과) ;
  • 천두성 (국립보건원 바이러스과) ;
  • 조해월 (국립보건원 바이러스과) ;
  • 장경립 (부산대학교 미생물학과)
  • Published : 2002.02.01
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Abstract

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.

굴 소비자의 보호를 위해서는 매우 민감하고 특이적으로 바이러스를 모니터링할 수 있는 신속진단법의 개발이 필수적이다. 굴 조직은 흔히 비교적 소량의 바이러스와 검출단계를 방해할 수 있는 다른 물질들을 함께 함유한다. 따라서 굴으로부터 바이러스의 검출에서 가장 중요한 과정은 시료의 가공단계이다. 본 연구에 의하면 한번의 sucrose 구배 초원심분리에 의하여 10%와 50% 사이에서 소량의 바이러스를 분리할 수 있음을 제시하였다. 우리는 두 종류의 primer 세트를 이용하여 HAV와 poliovirus를 동시에 굴 조직으로부터 검출할 수 있었다. 또한, 이 방법은 높은(>95%) 바이러스 회수율을 나타내었다. 이 방법은 24시간이내에 5 g의 굴조직으로부터 2 pfu의 HAV를 검출할 수 있을 정도로 신속하고 민감한 검사법이다.

Keywords

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