우결핵 병리조직으로부터의 우형결핵균의 분리 및 신속동정 방법

Detection of mycobacterium bovis in the pathological lesions of slaughtered dairy cattles using modified culture medium and polymerase chain reaction

Twenty-seven specimens were sampled from the pathological lesions of the thirteen dairy cattle slaughtered owing to the positive tuberculin reaction. The conventional smear microscopy and the culture on BM medium were compared with the polymerase chain reaction(PCR) technique for the detection of the causative organism of bovine tuberculosis in the sample specimens. On the basis of nucleotide sequence of IS1081 and mtp40 each pair of oligonucleotides was used as primers for the PCR amplification to target 300bp and 396bp respectively. The oligonucleotide primers from IS1081 prodcued positive signals for M. bovis in the amplified reactions while the primer mtp40 did not make PCR product because of its specificity to M. tuberculosis. Of the 27 sample specimens 19(70.4%) were detected as positive by both of the concentrated smear microscopy and the culture on BM media, but 6 specimens were confirmed as positive only by culture. Thus the total cultrue positive sample were 25(92.6%). PCR using IS1081 primers detected 26(96.3%) of these sample as positive for M. bovis. From one PCR negative but the cultrue positive sample M. nonchromogenicum was isolated. Results of the PCR and the culture showed that M. bovis was successfully detected by our suggested methods from the pathological lesions of dairy cattle slaughtered.