Medical and Pharmaceutical Biotechnology, The Center for Research and Applied Technology in Jalisco (CIATEJ), Av. Normalistas No. 800, Col. Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco, Mexico.
Public Health Laboratory of Jalisco, Guadalajara, Jalisco, Mexico.
J Glob Antimicrob Resist. 2017 Dec;11:90-97. doi: 10.1016/j.jgar.2017.07.004. Epub 2017 Jul 29.
The objectives of this study were to analyse the frequency of gene mutations associated with antitubercular drug resistance in clinical samples from the population of Jalisco State (Mexico) and to evaluate the genetic variability of Mycobacterium tuberculosis and multidrug-resistant (MDR) M. tuberculosis strains to describe the frequency of various families.
Clinical isolates of M. tuberculosis obtained from Jalisco State were analysed. Isolates were subjected to drug susceptibility testing, and mutations were characterised by sequencing, followed by genotyping using spoligotyping and mycobacterial interspersed repetitive units-variable-number of tandem repeats (MIRU-VNTR). Moreover, the prevalence of mutations was analysed by phylogenetic lineages.
Resistant strains were analysed by sequencing of katG, inhA and rpoB genes to determine the presence of mutations associated with isoniazid and rifampicin resistance. In MDR, monoresistant and polyresistant isolates, mutations were found in 17 (54.84%) of 31 strains. Spoligotyping identified six different strain lineages [T1 (25.40%), H3 (7.94%), MANU (4.76%), X1 (3.17%), EAI5 (1.59%) and LAM1 (1.59%)], with the remaining strains identified as orphans. In additional tree-based identification, a dendrogram of spoligotype patterns generated five different similarity clusters. When combining 24-loci MIRU-VNTR and spoligotyping approaches, the results shows that there is no cluster formation, indicating low transmission of the samples.
This study using spoligotyping and MIRU-VNTR showed that the analysed strains were not related to each other since no two identical strains were found. Families with the highest prevalence in the study were orphans followed by T family.
本研究的目的是分析来自哈利斯科州(墨西哥)人群的临床样本中与抗结核药物耐药相关的基因突变频率,并评估结核分枝杆菌和耐多药(MDR)结核分枝杆菌菌株的遗传变异性,以描述各种家族的频率。
分析了来自哈利斯科州的结核分枝杆菌临床分离株。对分离株进行药物敏感性测试,通过测序鉴定突变,并通过 spoligotyping 和分枝杆菌插入重复单元-可变数串联重复(MIRU-VNTR)进行基因分型。此外,通过系统发育谱系分析了突变的流行率。
通过测序 katG、inhA 和 rpoB 基因分析耐药菌株,以确定与异烟肼和利福平耐药相关的突变。在 MDR、单耐药和多耐药分离株中,在 31 株中有 17 株(54.84%)发现突变。 spoligotyping 鉴定出了 6 个不同的菌株谱系[T1(25.40%)、H3(7.94%)、MANU(4.76%)、X1(3.17%)、EAI5(1.59%)和 LAM1(1.59%)],其余菌株被鉴定为孤儿。在基于树的额外鉴定中, spoligotype 模式生成了 5 个不同的相似性聚类的树状图。当结合 24 位 MIRU-VNTR 和 spoligotyping 方法时,结果表明没有聚类形成,表明样本的传播性低。
本研究使用 spoligotyping 和 MIRU-VNTR 表明,分析的菌株彼此之间没有关系,因为没有发现两个相同的菌株。在研究中,流行率最高的家族是孤儿,其次是 T 家族。