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중합효소 연쇄반응을 이용한 돼지 내인성 레트로 바이러스의 검출과 분류

Detection and Classification of Porcine Endogenous Retroviruses by Polymerase Chain Reaction

  • 이동희 (건국대학교 동물생명과학대학 동물생명공학과) ;
  • 이정은 (건국대학교 동물생명과학대학 동물생명공학과) ;
  • 김훈미 (건국대학교 동물생명과학대학 동물생명공학과) ;
  • 김계웅 (공주대학교 산업과학대학 동물자원학과) ;
  • 박홍양 (건국대학교 동물생명과학대학 동물생명공학과) ;
  • 김영봉 (건국대학교 동물생명과학대학 동물생명공학과)
  • Lee, D.H. (Department of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University) ;
  • Lee, J.E. (Department of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University) ;
  • Kim, H.M. (Department of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University) ;
  • Kim, G.W. (Department of Animal Resources Science, College of Industrial Science, Kongju National University) ;
  • Park, H.Y. (Department of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University) ;
  • Kim, Young-Bong (Department of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University)
  • 발행 : 2007.06.30

초록

돼지의 내인성 레트로 바이러스인 PERV의 존재는 돼지의 무균 사육을 통해서도 제거할 수 없으며, 장기 이식 시 인수 공통 감염의 위험성을 내포하고 있으므로, 이종 간 장기 이식 시 면역 거부 반응과 더불어 해결해야 할 가장 큰 문제점 중의 하나이다. 본 연구에서는 국내에서 사육되고 있는 Berkshire, Duroc, Landrace, Yorkshire 종을 비롯하여 국내 미니 돼지 및 제주도 토종 흑돼지에서 PERV의 존재 유무 및 종류에 대하여 확인 및 검출하는 중합효소 연쇄반응(PCR) 기반 기법을 확립하였다. 사용된 모든 공시 돼지의 genomic DNA로부터 PERV- A 및 PERV-B가 모두 검출되었으며, PERV-C의 경우 음성대조구인 PK15 세포에서의 결과와 비교했을 때, Duroc종에서 매우 낮은 양의 PCR 산물이, 국내 미니돼지와 제주 토종 흑돼지에서는 상대적으로 높은 양의 PCR 산물이 검출되었다. 또한 PERV-A와 PERV-B의 경우 특이적 primer와 제한효소 BamHI을 이용한 PCR- RFLP를 통하여 확인할 수 있었다. 본 연구의 PERV 검출법은 이종 간 장기 이식 시 PERV에 대한 안전 검정 및 PERV의 분류 방법으로서 활용가능할 것으로 사료된다.

Pigs have been considered as an ideal source of donor organs because of their plentiful supply and their numerous anatomical and physiological similarities to the human in xenotransplantation. However, for the public health risks associated with the potential for porcine endogenous retrovirus(PERV) infection through xenograft from pig to human, the investigation of methods for elimination and/or control of PERV has been required. In this study we developed the detection and classification methods for PERV based on PCR using specific primers. PERV-A and PERV-B were found in all pigs including Berkshire, Duroc, Landrace, Yorkshire, miniature pig, and Korean native black pig from Jeju by PCR with type-specific primers for PERV. However, PERV-C was detected only from Duroc, miniature pig, and Korean native black pig from Jeju. PERV-A and PERV-B could be distinguished by PCR-RFLP with BamHI. These methods for PERV will be useful in rapid screening of safe organ for xenograft, furthermore, helpful in monitoring of PERV during and after xenotransplantation.

키워드

참고문헌

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