지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2011.1
- 수록면
- 321 - 331 (11page)
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초록· 키워드
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Mycobacterium bovis, a member of the M. tuberculosis complex (MTC), is the causative agent of bovine tuberculosis. Detection of M. bovis and M. tuberculosis using conventional culture- and biochemical-based assays is time-consuming. Therefore, a simple and sensitive molecular assay for rapid detection would be of great help in specific situations such as faster diagnosis of bovine tuberculosis (bTB) infection in the abattoirs. We developed a novel multiplex real-time PCR assay which was applied directly to biological samples with evidence of bTB and it was allowed to differentiate between M. bovis and M. tuberculosis. The primers and TaqMan probes were designed to target the IS1081 gene, the multi-copy insertion element in the MTC and the 12.7-kb fragment which presents in M. tuberculosis, not in the M. bovis genome. The assay was optimized and validated by testing 10 species of mycobacteria including M. bovis and M. tuberculosis, and 10 other bacterial species such as Escherichia coli, and cattle lymph nodes (n=113). The tests identified 96.4% (27/28) as M. bovis from the MTC-positive bTB samples using conventional PCR for specific insertion elements IS1081. And MTC-negative bTB samples (n=85) were tested using conventional PCR and the real-time PCR. When comparative analyses were conducted on all bovine samples, using conventional PCR as the gold standard, the relative accuracy of real-time PCR was 99.1%, the relative specificity was 100%, and the agreement quotient (kappa) was 0.976. The detection limits of the real-time PCR assays for M. bovis and M. tuberculosis genomic DNA were 10 fg and 0.1 pg per PCR reaction, respectively. Consequently, this multiplex real-time PCR assay is a useful diagnotic tool for the identification of MTC and differentiation of M. bovis and M. tuberculosis, as well as the epidemiologic surveillance of animals slaughtered in abattoir.
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참고문헌 (25)
참고문헌 신청
[학술지(정기
Bakshi CS / 2005 / Rapid differentiation of Mycobacterium bovis and Mycobacterium tuberculosis based on a 12.7-kb fragment by a single tube multiplex-PCR / Vet Microbiol 109
Barouni AS / 2004 / A pncA polymorphism to differentiate between Mycobacterium bovis and Mycobacterium tuberculosis / Mol Cell Probes 18 (3) : 167 ~ 170
Brosch R / 2002 / A new evolutionary scenario for the Mycobacterium tuberculosis complex / Proc Natl Acad Sci USA 99 : 3684 ~ 3689
Chen R / 2010 / Development of a fluorescent microsphere-based multiplex assay for simultaneous rapid detection of Mycobacterium tuberculosis complex and differentiation of M
Bakshi CS / 2005 / Rapid differentiation of Mycobacterium bovis and Mycobacterium tuberculosis based on a 12.7-kb fragment by a single tube multiplex-PCR / Vet Microbiol 109
Barouni AS / 2004 / A pncA polymorphism to differentiate between Mycobacterium bovis and Mycobacterium tuberculosis / Mol Cell Probes 18 (3) : 167 ~ 170
Brosch R / 2002 / A new evolutionary scenario for the Mycobacterium tuberculosis complex / Proc Natl Acad Sci USA 99 : 3684 ~ 3689
Chen R / 2010 / Development of a fluorescent microsphere-based multiplex assay for simultaneous rapid detection of Mycobacterium tuberculosis complex and differentiation of M
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