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Á¦¸ñ ³ª±Õ »ýÁ¸ Æò°¡¸¦ À§ÇÑ Propidium MonoazideÀ» ÀÌ¿ëÇÑ ½Ç½Ã°£ ÁßÇÕÈ¿¼Ò¿¬¼â¹ÝÀÀÀÇ ÀÇÀÇ
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³âµµ 2016 ±Ç 49
È£ 1 ¹øÈ£ 2
½ÃÀÛÆäÀÌÁö 13 ³¡ÆäÀÌÁö 22
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¿ä¾à 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 ¥ÄCT values (CT value in PMA-treated samples-CT value in non- PMA-treated samples). Real-time PCR was performed using QuantiTect SYBR¢ç 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 ¥ÄCT 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.
¡Ø Key Words :
Mycobacterium leprae , Propidium monoazide, Real-time PCR
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