评估一种新的检测瘰疬分枝杆菌菌株的real-time PCR方法

目的 建立针对非结核分枝杆菌中瘰疬分枝杆菌菌株的real-time PCR检测方法。方法 在我们的研究中,根据瘰疬分枝杆菌的SodA基因设计MGB探针和特定的引物,并用来检测模拟样品中的瘰疬分枝杆菌。结果 该方法 最低瘰疬分枝杆菌DNA检测浓度是10¹拷贝数,标准曲线显示阈值周期和SodA基因片段拷贝数之间的相关系数是0.973,斜率为-3.249,表现出良好的线性关系。此外, 模拟样品最低检测浓度是每毫升10¹个细菌,此外,其他9株菌为阴性结果 ,验证了良好的特异性。结论 该方法对于检测瘰疬分枝杆菌模拟标本显示很高的敏感性和特异性,可用于瘰疬分枝杆菌菌株的生态和流行病学监测。

Evaluation of a new real-time PCR assay for detection of Mycobacterium scrofulaceum strains

Mycobacterium scrofulaceum (M. scrofulaceum) is a slow-growing environmental and opportunistic atypical mycobacterium. In our study, MGB probes and specific primers were designed according to SodA gene of M. scrofulaceum, and the sensitive, specific and rapid real-time PCR assay for M. scrofulaceum was established, and was used to detect M. scrofulaceum in simulation samples. The minimum detectable concentration was 10¹ copies for M. scrofulaceum DNA. The standard curve showed correlation coefficient between threshold cycle and SodA gene fragment copy number was 0.973 and slope was -3.249 which showed a good linear relationship. In addition, the minimum detectable concentration was 10¹ Cells per milliliter for simulation sample. Also, the other nine strains showed negative results by the assay, which proved good specificity. This assay had high sensitivity and specificity for identification of M. scrofulaceum from the simulation specimens. The established real-time PCR should be useful for ecological and epidemiological surveillance of M. scrofulaceum strains.