Phenotypic and Genotypic Analysis of Multidrug-Resistant Mycobacterium tuberculosis Isolates from Sudanese Patients

Solima M Sabeel, M. Salih, M. Ali, S. El-Zaki, N. Abuzeid, Z. A. M. Elgadi, Hisham N. Altayb, A. M. A. Elegail, N. Ibrahim, B. K. Elamin
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引用次数: 26

Abstract

Background. Currently, mutations in rpoB, KatG, and rrs genes and inhA promoter were considered to be involved in conferring resistance to rifampicin, isoniazid, and streptomycin in Mycobacterium tuberculosis (MTB). Objective. The aims of this study were to detect the prevalence of first-line tuberculosis (TB) drug resistance among a group of previously treated and newly detected TB patients, to determine the association between prevalence of multidrug resistance (MDR) and demographic information (age and sex), to explain genes correlated with MDR Mycobacterium tuberculosis, and to characterize MTB via 16S ribosomal RNA (16S rRNA) analysis. Methods. A hundred MTB isolates from Sudanese pulmonary TB patients were included in the study. The proportional method of drug susceptibility test was carried out on Löwenstein-Jensen media. Multiplex PCR of rpoB and KatG genes and inhA promoter was conducted; then rrs genes were amplified by conventional PCR and were sequenced. The sequences of the PCR product were compared with known rrs gene sequences in the GenBank database by multiple sequence alignment tools. Result. The prevalence of MDR was 14.7% among old cases and 5.3% among newly diagnosed cases. Conclusion. Mutations in rrs could be considered as a diagnostic marker.
苏丹多药耐药结核分枝杆菌分离株表型和基因型分析
背景目前,rpoB、KatG和rrs基因以及inhA启动子的突变被认为与结核分枝杆菌(MTB)对利福平、异烟肼和链霉素的耐药性有关。客观的本研究的目的是检测一组既往治疗和新发现的结核病患者中一线结核病(TB)耐药性的患病率,确定多药耐药性(MDR)患病率与人口统计学信息(年龄和性别)之间的关系,解释与MDR结核分枝杆菌相关的基因,并通过16S核糖体RNA(16S rRNA)分析来表征MTB。方法。来自苏丹肺结核患者的100个MTB分离株被纳入该研究。在Löwenstein-Jensen培养基上进行了药敏试验的比例法。对rpoB和KatG基因及inhA启动子进行多重PCR;然后通过常规PCR扩增rrs基因并测序。通过多种序列比对工具将PCR产物的序列与GenBank数据库中的已知rrs基因序列进行比较。后果MDR的患病率在老年病例中为14.7%,在新诊断病例中为5.3%。结论rrs的突变可以被认为是一种诊断标记。
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