背景:将结核分枝杆菌(MTB)与非结核分枝杆菌(NTM)区分在患者的治疗过程中非常重要。根据美国胸科学会指南(ATS),应在适当的治疗和患者管理的物种水平上鉴定NTM临床分离株。本研究旨在通过评估rpoB来鉴定NTM临床分离株,ssrA,tuf,atpe,ku,和dnaK基因,并使用多位点序列分析(MLSA)将六个基因连接起来。
方法:96株分枝杆菌,包括86个NTM和10个MTB分离株,纳入了转诊到某些结核病参考中心的所有患者。通过PCR扩增rpoB对所有分离物进行评估,ssrA,tuf,ku,atpe,dnaK基因和MLSA.
结果:在96个分离株中,91(94.8%),87(90.6%),72(75%),通过rpoB将84(87.5%)和79(82.3%)区分为物种水平,tuf,ssrA,dnaK和atpE基因,分别。ku基因能够将86个NTM分离株中的69个(80.2%)分离株鉴定到物种水平。我们可以通过MLSA将100%的分离株鉴定到物种水平。
结论:本研究中使用的PCR靶标均不能完全区分所有物种。用于连接六个基因的MLSA技术可以增加临床分枝杆菌分离株的鉴定,并且所有16个物种均具有良好的分化。
BACKGROUND: Differentiating Mycobacterium tuberculosis (MTB) from nontuberculous mycobacteria (NTM) is very important in the treatment process of patients. According to the American Thoracic Society guideline (ATS), NTM clinical isolates should be identified at the species level proper treatment and patient management. This study aimed to identify NTM clinical isolates by evaluationg rpoB, ssrA,
tuf, atpE, ku, and dnaK genes, and use multilocus sequence analysis (MLSA) to concatenate the six genes.
METHODS: Ninety-six Mycobacterium isolates, including 86 NTM and 10 MTB isolates, from all the patients referred to the certain TB Reference Centres were included. All isolates were evaluated by PCR amplification of rpoB, ssrA,
tuf, ku, atpE, and dnaK genes and MLSA.
RESULTS: Out of 96 isolates, 91 (94.8%), 87 (90.6%), 72 (75%), 84 (87.5%) and 79 (82.3%) were differentiated to the species level by rpoB,
tuf, ssrA, dnaK and atpE genes, respectively. The ku gene was able to identify 69 (80.2%) isolates of the 86 NTM isolates to the species level. We could identify 100% of the isolates to the species level by MLSA.
CONCLUSIONS: None of the PCR targets used in this study were able to completely differentiate all species. The MLSA technique used to concatenate the six genes could increase the identification of clinical Mycobacterium isolates and all 16 species were well-differentiated.