PDF(Pubmed)
Abstract：
The Non-tuberculous mycobacterial (NTM) isolates should be distinguished from tuberculosis and identified at the species level for choosing an appropriate treatment plan. In this study, two molecular methods were used to differentiate NTM species, including a new designed High Resolution Melting (HRM) and Multilocus Sequence Analysis (MLSA). Seventy-five mycobacterial isolates were evaluated by sequencing four genes ( MLSA) and a HRM assay specifically targeting atpE was designed to rapidly and accurately identify and differentiate mycobacterium species. Out of 70 NTM isolates, 66 (94.3%), 65 (92.9%), 65 (92.9%) and 64 (91.4%) isolates were identified to the species level by PCR of atpE, tuf, rpoB and dnaK genes. We could identify 100% of the isolates to the species level (14 different species) by MLSA. By using HRM assay, all NTM isolates were identified and classified into eight groups, in addition, Mycobacterium tuberculosis and Nocardia were also detected simultaneously. The MLSA technique was able to differentiate all 14 species of NTM isolates. According to the results, the HRM assay is a rapid and beneficial method for identifying NTM, M. tuberculosis (MTB), and Nocardia isolates without sequencing.
摘要：
应将非结核分枝杆菌（NTM）分离株与结核病区分开来，并在物种水平上进行鉴定，以选择合适的治疗方案。在这项研究中,两种分子方法被用来区分NTM物种，包括新设计的高分辨率熔解(HRM)和多位点序列分析(MLSA)。通过对四个基因（MLSA）进行测序来评估75个分枝杆菌分离株，并设计了专门针对atpE的HRM测定法，以快速，准确地鉴定和区分分枝杆菌物种。在70个NTM分离株中，66（94.3%），65（92.9%），通过atpE的PCR鉴定了65个（92.9％）和64个（91.4％）分离株，tuf,rpoB和dnaK基因。我们可以通过MLSA将100％的分离株鉴定到物种水平（14种不同的物种）。通过使用HRM分析，所有NTM分离株被鉴定并分为八组，此外,同时检测到结核分枝杆菌和诺卡氏菌。MLSA技术能够区分所有14种NTM分离株。根据结果,HRM测定法是一种快速和有益的鉴定NTM的方法，结核分枝杆菌（MTB），和没有测序的诺卡氏菌分离株。
