Umar F, Hatta M, Husain D R, Natzir R, Dwiyanti R, Junita A R, Primaguna M R
Makassar Medical State Laboratory, Indonesian Ministry of Health, Makassar, Indonesia.
Post Graduate Programme of Medical Science, Faculty of Medicine, University of Hasanuddin, Makassar, Indonesia.
New Microbes New Infect. 2019 Oct 8;32:100609. doi: 10.1016/j.nmni.2019.100609. eCollection 2019 Nov.
Efflux pumps are transmembrane proteins that vigorously participate in extruding a wide range of substrates, including drugs, outside the bacterial cell. We aimed to investigate the mRNA expression level of the efflux pump gene in isolated from individuals with tuberculosis who received drug therapy, at the 1st, 3rd and 5th months, and newly diagnosed patients with tuberculosis who will receive drug therapy (0 month). The study was a multiple cross-sectional longitudinal design-50 different isolates and a reference strain H37Rv were subcultured in LJ medium and confirmed by multiplex PCR for identification of and collected for RNA extraction. Total bacterial mRNA was analysed using real-time quantitative PCR to evaluate mRNA quantification gene expression. There were differences in the level of 250 mRNA expression between sensitive ( = 11) and resistant ( = 40) groups of 5.961 ± 0.414 and 10.192 ± 1.978, respectively (fold changes; p < 0.05). There were significant differences of expression level among -resistant groups (p < 0.05) specifically 7.573 ± 0.424 for 0-month drug therapy ( = 10), 9.438 ± 0.644 for 1st month drug therapy ( = 10), 11.057 ± 0.262 for 3rd month drug therapy ( = 10) and 12.701 ± 0.460 for 5th month drug therapy ( = 10). We assume that the extent of gene expression in clinical isolates is a result of the exposure to antimicrobials during treatment, which affect the basic expression of the efflux pump gene.
外排泵是一种跨膜蛋白，积极参与将包括药物在内的多种底物排出细菌细胞外。我们旨在研究从接受药物治疗的结核病患者中分离出的结核分枝杆菌外排泵基因在第1、3和5个月时的mRNA表达水平，以及新诊断的即将接受药物治疗（0个月）的结核病患者的情况。该研究采用多横断面纵向设计，将50株不同的分离株和一株参考菌株H37Rv在LJ培养基中传代培养，并通过多重PCR进行鉴定，然后收集用于RNA提取。使用实时定量PCR分析总细菌mRNA，以评估mRNA定量基因表达。敏感组（n = 11）和耐药组（n = 40）的结核分枝杆菌mRNA表达水平存在差异，分别为5.961 ± 0.414和10.192 ± 1.978（倍数变化；p < 0.05）。耐结核分枝杆菌组之间的表达水平存在显著差异（p < 0.05），具体而言，0个月药物治疗组（n = 10）为7.573 ± 0.424，1个月药物治疗组（n = 10）为9.438 ± 0.644，3个月药物治疗组（n = 10）为11.057 ± 0.262，5个月药物治疗组（n = 10）为12.701 ± 0.460。我们认为临床分离株中结核分枝杆菌基因的表达程度是治疗期间接触抗菌药物的结果，这会影响外排泵结核分枝杆菌基因的基础表达。
