Korean Journal of Veterinary Research 2001;41(4):535-542.
Multiplex PCR for differential diagnosis of Mycobacterium species from bovine clinical samples
Yong-hwan Kim1, MH Al-Haddawi2, Ho-seong Cho2, Sung-kwi Kang2, Kyoung-oh Cho2, Hyung-seon Park2, Bong-joo Lee2, Nam-yong Park2
1Gwangju Metropolitan City Institute of Health and Environment
2College of Veterinary Medicine, Chonnam National University
소의 임상병리 가검물에서 Mycobacterium species 감별진단을 위한 multiplex PCR 기법
김용환1, 2, 조호성2, 강성귀2, 조경오2, 박형선2, 이봉주2, 박남용2
1광주광역시보건환경연구원
2전남대학교 수의과대학
Abstract
A multiplex PCR technique was developed for detecting specifically each Mycobacterium bovis, M. tuberculosis, M. avium and M. avium subsp, paratuberculosis, respectively, using clinical samples of field cattle. To apply this novel technique to clinical specimens, blood sample was obtained from live cows comprising 11 intradermal tuberculin test (ITT)-positive and 17 ITT-negative and tested by multiplex PCR. Positive results were obtained from 15 cows by the multiplex PCR, showing that 4 (23.5%) of the 17 ITT-negative cows were multiplex PCR positive. The multiplex PCR results also showed that among the 15 positive cows, 7 (46.7%) were infected with M. bovis, 1 (6.7%) with M. tuberculosis and 7 (46.7%) with M. avium. The sensitivity and specificity of multiplex PCR in comparison with those of ITT were 100% and 76.5%. The correlation between the multiplex PCR and ITT assays with blood samples was considered excellent, 85.7% agreement and ${kappa}=0.72$. The results obtained, using reference mycobacterial strains and typed clinical samples, show that the multiplex PCR method may be a rapid, sensitive, and specific tool for the differential identification of various mycobacterial strains in a single-step assay. Therefore, multiplex PCR assay is a useful tool for early diagnosis of tuberculosis in live cattle and to identify the species or complex of mycobacterium from clinical samples.
Key Words: Bovine, Clinical samples, Intradermal tuberculin test, Multiplex PCR, Mycobacterium


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