Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
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Detection of Mycobacterium bovis in the lymph node of tuberculin positive cattle by guanidium isothiocyanate/silica DNA extraction and polymerase chain reaction

  • Cho, Yun-Sang (Bacteriology and Parasitology Division, Animal Disease Research Department, National Veterinary Research and Quarantine Service) ;
  • Jung, Suk-Chan (Laboratory of Infectious Diseases, College of, Veterinary Medicine and the School of Agricultural Biotechnology, Seoul National University) ;
  • Yoo, Han-Sang (Laboratory of Infectious Diseases, College of, Veterinary Medicine and the School of Agricultural Biotechnology, Seoul National University) ;
  • Kim, Jong-Man (Bacteriology and Parasitology Division, Animal Disease Research Department, National Veterinary Research and Quarantine Service)
  • Published : 2007.06.30
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Abstract

Tuberculin positive cattle without gross tubercle lesions should be confirmed by the bacteriological examination to determine the state of the infection. To overcome the time-consuming and laborious identification by culture and biochemical tests, polymerase chain reaction (PCR) has been used to identify Mycobacterium bovis. Due to various lipids in the cell wall of Mycobacterium spp, novel methods of DNA extraction from Mycobacterium spp have been developed. In this study, a newly developed guanidium isothiocyanate/silica DNA extraction method was directly applied to specimens from the tuberculin positive cattle. DNAs were directly extracted from the lymph nodes and the major polymorphic tandem repeat (MPTR) and mycobacterial protein of BCG 70 (MPB70) were amplified using PCR. The DNA extraction method using guanidium isothiocyanate/silica was efficient and safe, and the MPTR and MPB70 primers were specific to M bovis. Therefore, MPTR and MPB70 PCRs will be useful for the detection of M bovis in the lymph node from skin-test positive cattle.

Keywords

References

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