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제목 |
나균 생존 평가를 위한 Propidium Monoazide을 이용한 실시간 중합효소연쇄반응의 의의 |
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저자 |
김종필, 김연실, 김찬우 |
소속 |
한국한센복지협회 연구원 |
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년도 |
2016 |
권 |
49 |
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호 |
1 |
번호 |
2 |
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시작페이지 |
13 |
끝페이지 |
22 |
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첨부 |
2016년 나학회지2.pdf |
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요약 |
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. Object : The author evaluated whether PMA real-time PCR is suitable for the viablity of Mycobacterium
leprae ( M. leprae) in specimens of cultivation in mouse foot pads. Methods : A total of 55 diluted suspensions from mouse foot pads were quadruplicated and subjected to PMA treatment and/or heat inactivation, and were also tested to compare the ΔCT values (CT value in PMA-treated samples-CT value in non- PMA-treated samples). Real-time PCR was performed using QuantiTect SYBR® Green PCR Kits(Qiagen, USA), and the CT value changes after PMA treatment were compared between PMA treatment and/or heat inactivation groups. Results : The increase in the CT value after PMA treatment was significant in heat inactivated group(4.26) and non-heat inactivated group(1.12)(both P = 0.000). In the ROC curve analysis, the cutoff ΔCT value for maximum sensitivity (100%) and specificity (97.1%) for differentiating dead from live cells was 2.41 Conclusions : PMA real-time PCR is a useful approach for evaluating viablity of M. leprae.
※ Key Words :
Mycobacterium leprae , Propidium monoazide, Real-time PCR |
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내용 |
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