Rapid Diagnosis of Iridovirus Infection by Polymerase Chain Reaction

Polymerase Chain Reaction(PCR)을 이용한 Iridovirus의 검색

  • Cha, Seung-Ju (Department of Biological Science, University of Ulsan) ;
  • Do, Jeong-Wan (Department of Biological Science, University of Ulsan) ;
  • Kim, Hyun-Ju (Department of Biological Science, University of Ulsan) ;
  • Cho, Wha-Ja (Department of Biological Science, University of Ulsan) ;
  • Mun, Chang-Hoon (Department of Biological Science, University of Ulsan) ;
  • Park, Jeong-Min (Department of Biological Science, University of Ulsan) ;
  • Park, Myoung-Ae (Pathology Division, National Fisheries Research and Development Agency) ;
  • Kim, Su-Mi (Pathology Division, National Fisheries Research and Development Agency) ;
  • Sohn, Sang-Gyu (Pathology Division, National Fisheries Research and Development Agency) ;
  • Bang, Jong-Deuk (Pathology Division, National Fisheries Research and Development Agency) ;
  • Park, Jeong-Woo (Department of Biological Science, University of Ulsan)
  • Published : 1998.05.30

Abstract

For rapid detection of iridovirus infection, a PCR-based diagnostic method was developed. The genomic DNA from mortality-associated iridovirus was cloned into pUC19 vector. The nucleotide sequences of these clones were compared with sequences of other genes from EMBL/GenBank databank. Based on the nucleotide sequences, PCR primers were prepared and used for PCR. The DNA amplification did not occur from the normal fish cells. In contrast, DNA was amplified from the iridovirus-infected fish cells and purified iridovirus. These results suggest that mortality-associated iridovirus can be detected from virus-infected cells within short time and this PCR-based diagnostic system provides a simple and accurate method for detecting the presence of iridovirus infection.

PCR을 사용하여 양식 해산어에 iridovirus 감염 여부를 신속히 진단하고자 하였다. 먼저 폐사를 유발하는 iridovirus의 genomic DNA를 pUC19 vector에 cloning한 후 이 clone들의 염기 서열을 GenBank의 염기 서열과 비교 분석하였다. 이들의 염기 서열을 기초로 하여 PCR primer를 제조한 후 PCR을 수행하였다. 정상적인 어류 세포에서는 DNA의 증폭이 일어나지 않았으나 iridovirus에 감염된 세포와 순수 분리된 iridovirus에서는 DNA의 증폭이 일어났다. 이로부터 짧은 시간 내에 폐사를 유발하는 iridovirus를 신속히 진단할 수 있음이 확인되었으며 이러한 PCR을 이용한 진단은 iridovirus의 감염을 확인할 수 있는 간단하고도 정확한 방법을 제공해준다.

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