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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제25권 제5호
- 발행연도
- 2015.1
- 수록면
- 745 - 752 (8page)
이용수
초록· 키워드
오류제보하기
Tuberculosis (TB) is the most common mycobacterial infection in developing countries, requiring a rapid, accurate, and well-differentiated detection/diagnosis. For the rapid detection and discrimination of Mycobacterium tuberculosis complex (MTC) from nontuberculous mycobacteria (NTM), a novel, simple, and primer-combined single-step multiplex PCR using three primer pairs (6110F-6110R, 1081F-1081R, and 23SF-23SR; annealing on each of IS6110, IS1081, and 23S rDNA targets), hereafter referred to as a triplex PCR, has been developed and evaluated. The expected product for IS6110 is 416 bp, for IS1081 is 300 bp, and for 23S rDNA is 206 bp by single PCR, which was used to verify the specificity of primers and the identity of MTC using DNA extracted from the M. tuberculosis H37Rv reference strain (ATCC, USA) and other mycobacteria other than tuberculosis (MOTT) templates. The triplex PCR assay showed 100% specificity and 96% sensitivity; the limit of detection for mycobacteria was ~100 fg; and it failed to amplify any target from DNA of MOTT (50 samples tested). Of 307 blinded clinical samples, overall 205 positive M. tuberculosis samples were detected by single PCR, 142 by conventional culture, and 90 by AFB smear methods. Remarkably, the triplex PCR could subsequently detect 55 positive M. tuberculosis from 165 culture-negative and 115 from 217 AFB smear-negative samples. The triplex PCR, targeting three regions in the M. tuberculosis genome, has proved to be an efficient tool for increasing positive detection/ discrimination of this bacterium from clinical samples.
목차
등록된 정보가 없습니다.
참고문헌 (22)
참고문헌 신청
Alcaide F / 2000 / Evaluation of the BACTEC MGIT 960 and the MB/BacT systems for recovery of mycobacteria from clinical specimens and for species identification by DNA AccuPr
Britten RJ / 2005 / The majority of human genes have regions repeated in other human genes / Proc. Natl. Acad. Sci. USA 102 : 5466 ~ 5470
Cegielski JP / 1997 / Comparison of PCR, culture, and histopathology for diagnosis of tuberculous pericarditis / J. Clin. Microbiol 35 : 3254 ~ 3257
Collins DM / 1991 / Identification of an insertion sequence, IS1081, in Mycobacterium bovis / FEMS Microbiol. Lett 67 : 11 ~ 15
Collins DM / 1993 / DNA fingerprinting of Mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements IS1081 and IS6110 / J. Clin.
Britten RJ / 2005 / The majority of human genes have regions repeated in other human genes / Proc. Natl. Acad. Sci. USA 102 : 5466 ~ 5470
Cegielski JP / 1997 / Comparison of PCR, culture, and histopathology for diagnosis of tuberculous pericarditis / J. Clin. Microbiol 35 : 3254 ~ 3257
Collins DM / 1991 / Identification of an insertion sequence, IS1081, in Mycobacterium bovis / FEMS Microbiol. Lett 67 : 11 ~ 15
Collins DM / 1993 / DNA fingerprinting of Mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements IS1081 and IS6110 / J. Clin.
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