Detection of porcine encephalomyocarditis virus by in situ hybridization

In situ hybridization에 의한 돼지 뇌심근염 바이러스의 검출

  • Oh, Sang-hyeon (College of Veterinary Medicine, Chonnam National University) ;
  • Park, Nam-yong (College of Veterinary Medicine, Chonnam National University) ;
  • Chung, Ci-young (College of Veterinary Medicine, Chonnam National University) ;
  • Cho, Kyoung-oh (College of Veterinary Medicine, Chonnam National University) ;
  • Lee, Bong-joo (College of Veterinary Medicine, Chonnam National University) ;
  • Park, Young-seok (College of Veterinary Medicine, Chonnam National University) ;
  • Park, Hyung-seon (College of Veterinary Medicine, Chonnam National University)
  • Received : 1999.02.18
  • Published : 1999.03.22

Abstract

The purpose of this study was to establish a rapid, reliable diagnostic method detecting Encephalomyocarditis virus(EMCV) RNA in formalin-fixed, paraffin-embedded tissues of EMCV naturally infected pigs by cDNA probe of EMC $K_3$, the EMCV strain isolated from Korea. Using a biotin-labelled nick translated probe for the cDNA marker. We made up for some defects of radiolabeled method. In sits hybridization(ISH) technique, differently from the other nucleic acid hybridization methods, is able to detect the virus genome specifically in the state of the intact shapes of cells and/or tissues. We succeeded in performing the experiment to detect the EMCV within 1~2 hours using the $MicroProbe^{TM}$ capaillary action system. In this study, we observed highly specific positive signals of red color by staining the paraffin-embedded tissue sections of naturally EMCV-infected pig organs or tissues, including brain, heart, kidney and lacrimal gland with the Fast Red TR salt/Naphtol phosphate chromogen. The results suggested that this ISH method is considered as a highly sensitive and reliable tool for molecular biologic diagnosis of the EMC viral disease.