Umar Faiqah F, Husain Dirayah R, Hatta Mochammad M, Natzir Rosdiana R, Sjahril Rizalinda S, Dwiyanti Ressy R, Junita Ade R, Primaguna Muhammad R
Post Graduate of Medical Science, Faculty of Medicine, University of Hasanuddin, Makassar, Indonesia.
Department of Biology, Faculty of Science, University of Hasanuddin, Makassar, Indonesia.
J Taibah Univ Med Sci. 2020 Feb 7;15(1):54-58. doi: 10.1016/j.jtumed.2019.12.003. eCollection 2020 Feb.
This study aimed to determine molecular characteristics of , and genes in isolated from a cohort of Indonesian patients with tuberculosis.
Fifty isolates of were analysed by testing (DST) for susceptibility to first- and second-line drugs using the proportional method in a liquid medium. The genomic material was extracted to perform multiplex polymerase chain reaction (PCR) for identification and gene sequencing of , and .
Approximately 80% (40/50) of the mutations that were detected outside the hot-spot region (S450L, H445D, D435V, S441L, I491F, and Q432P) conferred rifampicin-resistance on . Approximately 11.42% (4/35) of isolates with S315T mutation in led to rifampicin-resistance instead of isoniazid-resistance. The mutation in gene was found at various locations (P280P, G279R, E340Q, T271I, E340*stop codon, R373G, and S315N). Streptomycin-resistance was detected in 42% (21/50) of the strains, but only two strains had gene mutations (G878A and/or S514R). Approximately 14% (7/50) of isolates were kanamycin- and capreomycin-resistant but did not harbour mutations in the gene, while 80% (40/50) of the strains had mutations in the quinolone-resistance determining region (QRDR) of the gene (S95T, D94V, A90V, and S91P) including the pan-susceptible strain.
Of the 50 strains analysed, most of the mutations in the gene associated with rifampicin-resistance were also detected in the and genes. Molecular characterisation using DNA sequencing techniques is a highly sensitive approach for detecting mutations.
本研究旨在确定从一组印度尼西亚结核病患者中分离出的结核分枝杆菌中rpoB、katG和rrs基因的分子特征。
采用液体培养基中的比例法对50株结核分枝杆菌进行一线和二线药物敏感性检测（DST）。提取基因组材料以进行多重聚合酶链反应（PCR），用于rpoB、katG和rrs的鉴定及基因测序。
在热点区域（S450L、H445D、D435V、S441L、I491F和Q432P）之外检测到的约80%（40/50）的rpoB突变赋予了结核分枝杆菌对利福平的抗性。katG中S315T突变的约11.42%（4/35）分离株导致对利福平耐药而非对异烟肼耐药。rrs基因突变出现在不同位置（P280P、G279R、E340Q、T271I、E340*终止密码子、R373G和S315N）。42%（21/50）的菌株检测到对链霉素耐药，但只有两株有rrs基因突变（G878A和/或S514R）。约14%（7/50）的结核分枝杆菌分离株对卡那霉素和卷曲霉素耐药，但rrs基因未发生突变，而80%（40/50）的菌株在gyrA基因的喹诺酮耐药决定区（QRDR）有突变（S95T、D94V、A90V和S91P），包括全敏感菌株。
在分析的50株菌株中，与利福平耐药相关的rpoB基因中的大多数突变在katG和rrs基因中也被检测到。使用DNA测序技术进行分子特征分析是检测突变的一种高度灵敏的方法。
