• Title, Summary, Keyword: radiolabeled in situ hybridization

Search Result 2, Processing Time 0.032 seconds

Identification of Aujeszky's disease virus by in situ hybridization (In situ hybridization 조직화학법을 이용한 오제스키병 바이러스 동정)

  • Kim, Soon-bok;Sur, Jung-hang;Moon, Oun-kyung
    • Korean Journal of Veterinary Research
    • /
    • v.34 no.2
    • /
    • pp.327-333
    • /
    • 1994
  • The purpose of this study was to establish a rapid diagnostic method detecting Aujeszky's disease virus (ADV) DNA in the cultured cell monolayers (PK-15) and tissue sections of ADV(NYJ-1-87)-infected rats and pigs by in situ hybridization(ISH). Detection of specific ADV-DNA in infected cells was conducted by radiolabeled ISH method using $^{32}P-labeled $ DNA probe (BamH1 7 fragment) which contains a 6.3 Kb ADV-DNA insert. Where ADV-DNA was detected by radiolabeled ISH, the deposition of black photographic grains occurred in the nuclei and the cytoplasms of ADV-infected cells. Positive hybridization signal was often observed in the spinal trigerminal nucleus of the pons, the nucleus of the trigerminal ganglion neuron and the epithelial cells of tonsillar crypts. The results suggested that ISH is considered as a highly sensitive and reliable tool for confirmative diagnosis of this viral disease.

  • PDF

Detection of porcine encephalomyocarditis virus by in situ hybridization (In situ hybridization에 의한 돼지 뇌심근염 바이러스의 검출)

  • Oh, Sang-hyeon;Park, Nam-yong;Chung, Ci-young;Cho, Kyoung-oh;Lee, Bong-joo;Park, Young-seok;Park, Hyung-seon
    • Korean Journal of Veterinary Research
    • /
    • v.39 no.1
    • /
    • pp.148-158
    • /
    • 1999
  • 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.

  • PDF