지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2014.1
- 수록면
- 203 - 209 (7page)
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초록· 키워드
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Background: Conventional acid-fast bacilli (AFB) staining cannot differentiate viable from dead cells. Propidium monoazide (PMA) is a photoreactive DNA-binding dye that inhibits PCR amplification by DNA modification. We evaluated whether PMA real-time PCR is suit- able for the early detection of viable Mycobacterium tuberculosis (MTB) in clinical respira- tory specimens.
Methods: A total of 15 diluted suspensions from 5 clinical MTB isolates were quadrupli- cated and subjected to PMA treatment and/or heat inactivation. Eighty-three AFB-positive sputum samples were also tested to compare the ΔCγ T values ( Cγ T value in PMA-treated sputum samples− Cγ T value in non-PMA-treated sputum samples) between culture-positive and culture-negative specimens. Real-time PCR was performed using Anyplex MTB/NTM Real-Time Detection (Seegene, Korea), and the CγT value changes after PMA treatment were compared between culture-positive and culture-negative groups.
Results: In MTB suspensions, the increase in the CγT value after PMA treatment was sig- nificant in dead cells ( P =0.0001) but not in live cells ( P =0.1070). In 14 culture-negative sputum samples, the median ΔCγT value was 5.3 (95% confidence interval [CI], 4.1-8.2; P <0.0001), whereas that in 69 culture-positive sputum samples was 1.1 (95% CI, 0.7- 2.0). In the ROC curve analysis, the cutoff ΔCγT value for maximum sensitivity (89.9%) and specificity (85.7%) for differentiating dead from live cells was 3.4.
Conclusions: PMA real-time PCR is a useful approach for differentiating dead from live bacilli in AFB smear-positive sputum samples.
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참고문헌 (18)
참고문헌 신청
Blumberg HM / 2003 / American Thoracic Society/Centers for Disease Control and Prevention/Infectious Diseases Society of America: treatment of tuberculosis / Am J Respir Crit
Brescia CC / 2009 / Cryptosporidium propidium monoazide-PCR, a molecular biology-based technique for genotyping of viable Cryptosporidium oocysts / Appl Environ Microbiol 75
Causse M / 2011 / Comparison of two molecular methods for rapid diagnosis of extrapulmonary tuberculosis / J Clin Microbiol 49 : 3065 ~ 3067
Drobniewski FA / 2003 / Modern laboratory diagnosis of tuberculosis / Lancet Infect Dis 3 : 141 ~ 147
Fittipaldi M / 2012 / Progress in understanding preferential detection of live cells using viability dyes in combination with DNA amplification / J Microbiol Methods 91 : 276
Brescia CC / 2009 / Cryptosporidium propidium monoazide-PCR, a molecular biology-based technique for genotyping of viable Cryptosporidium oocysts / Appl Environ Microbiol 75
Causse M / 2011 / Comparison of two molecular methods for rapid diagnosis of extrapulmonary tuberculosis / J Clin Microbiol 49 : 3065 ~ 3067
Drobniewski FA / 2003 / Modern laboratory diagnosis of tuberculosis / Lancet Infect Dis 3 : 141 ~ 147
Fittipaldi M / 2012 / Progress in understanding preferential detection of live cells using viability dyes in combination with DNA amplification / J Microbiol Methods 91 : 276
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