Medical Technology Program, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand; Centre for Research and Development of Medical Diagnostic Laboratories (CMDL), Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.
Health Intervention and Technology Assessment Program (HITAP), Ministry of Public Health, Nonthaburi, Thailand.
Infect Genet Evol. 2020 Nov;85:104449. doi: 10.1016/j.meegid.2020.104449. Epub 2020 Jul 1.
Tuberculosis, caused by Mycobacterium tuberculosis (MTB) infection, remains a global health problem with increased concerns due to drug-resistant tuberculosis. However, molecular genotyping profiles may give insight of the transmission of TB in a particular region. The present study aimed to characterize the genetic diversity of drug-resistant MTB and evaluate primer sets applied for the epidemiological study of circulating MTB in Northeastern Thailand. A total of 92 MTB isolates, resistant to rifampicin and/or isoniazid, were collected from the Office of Disease Prevention and Control between 2013 and 2016. All isolates were genotyped by 24-locus MIRU-VNTR typing combined with spoligotyping. We also analyzed the distributions of drug susceptibility pattern and demographic data among different genotypes. In comparison with different loci sets, discriminatory power based on 12, 15, 24 standard primers were investigated. Eighty-six particular profiles were found; among the patterns, two clusters were produced in 8 strains. East African Indians (EAI) were the most prevalent strains (33 isolates, 35.87%) followed by Beijing (30 isolates, 32.61%), with 23 unknown isolates strains also found. The HGDI based on combination of 24 loci analysis and spoligotyping was 0.9962. The number of tandem repeat generated was highly discriminant (HGDI>0.6) at locus 580 (0.66), 960 (0.67), 2163b (0.73), 2165 (0.62), 2461 (0.68) 3690 (0.73) and 4052 (0.79), respectively. In contrast, the diversity at locus 154 and 2059 was not revealed. The results emphasized that 24-locus MIRU-VNTR and spoligotyping could be useful for epidemiological surveillance of drug-resistant MTB in this region. At a given allelic diversity, 7 primer sets containing MIRU04, MIRU10, QUB2163b, ETRA, ETRB, Mtub39 and QUB26 may be considered for screening the VNTR patterns. In addition, this study gathered both demographics and genotypic data within the same investigation for further tuberculosis prevention and control.
结核病是由结核分枝杆菌（MTB）感染引起的，由于耐多药结核病的出现，它仍然是一个全球性的健康问题。然而，分子基因分型谱可能可以深入了解特定地区结核病的传播情况。本研究旨在描述耐多药 MTB 的遗传多样性，并评估在泰国东北部用于流行性病学研究的引物集。从 2013 年到 2016 年，在疾病预防控制办公室收集了 92 株耐利福平（rifampicin）和/或异烟肼（isoniazid）的 MTB 分离株。所有分离株均通过 24 个基因座 MIRU-VNTR 分型结合 spoligotyping 进行基因分型。我们还分析了不同基因型之间的药物敏感性模式和人口统计学数据的分布。与不同的基因座集相比，我们研究了基于 12、15、24 个标准引物的区分力。发现了 86 种特定的模式；在这些模式中，有 8 株产生了 2 个聚类。东非印度人（EAI）是最常见的菌株（33 株，35.87%），其次是北京（30 株，32.61%），还发现了 23 株未知菌株。基于 24 个基因座分析和 spoligotyping 组合的 HGDI 为 0.9962。生成的串联重复数高度具有区分力（HGDI>0.6），在 580（0.66）、960（0.67）、2163b（0.73）、2165（0.62）、2461（0.68）、3690（0.73）和 4052（0.79）基因座上。相比之下，154 和 2059 基因座的多样性未被揭示。结果强调，24 个基因座 MIRU-VNTR 和 spoligotyping 可用于该地区耐多药 MTB 的流行病学监测。在给定的等位基因多样性下，7 个包含 MIRU04、MIRU10、QUB2163b、ETRA、ETRB、Mtub39 和 QUB26 的引物集可用于筛选 VNTR 模式。此外，本研究在同一调查中收集了人口统计学和基因型数据，以进一步进行结核病的预防和控制。
