[临床结核分枝杆菌菌株embB基因306密码子突变的分子遗传学检测]。

M V Afanas'ev, A D Borovskaia, E N Il'ina, T G Smirnova, E E Larionova, A V Kuz'min, S N Andreevskaia, L N Chernousova, V M Govorun
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引用次数: 0

摘要

本研究共使用结核分枝杆菌254株。其中对乙胺丁醇(EMB)耐药菌株183株,对EMB敏感的多药耐药菌株13株,对利福平(RIF)、异烟肼(INZ)和EMB敏感的菌株39株。对所有菌株的embB306、rpoB和katG/inhA启动子3个基因位点的遗传变化进行了分析,这3个基因位点分别与EMB、RIF和INZ抗性的形成有关。结核分枝杆菌菌株来自生活在俄罗斯联邦中部地区的肺结核患者。绝对浓度试验显示对RIF、INZ和EMB的抗性。测定了EMB对各菌株的抑菌浓度(IC)。上述基因座的遗传变化通过mini-sequencing估计,然后用质谱法记录MALDI-TOF产物。emb耐药菌株中embB306的突变频率相对较低(约为41.5%)。密码子306仅在EMB IC >或= 2 mg/L的菌株中检测到突变。embB306突变频率与多药耐药表型之间存在统计学上显著的关联。将这些突变与传统的多药耐药遗传标记相结合,可用于更有效地检测多药耐药菌株。
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[Detection of mutations in codon 306 of the embB gene for molecular genetic characterization of clinical Mycobacterium tuberculosis strains].

A total of 254 Mycobacterium tuberculosis strains were used in the study. Among them, there were 183 ethambutol (EMB)-resistant strains, 13 multidrug resistant ones, but EMB-sensitive, and 39 strains sensitive to rifampicin (RIF), isoniazid (INZ), and EMB. All the strains were analyzed for genetic changes in three loci: embB306, rpoB, and katG/inhA promoter, which were associated with the formation of resistance to EMB, RIF, and INZ, respectively. The Mycobacterium tuberculosis strains were obtained from pulmonary tuberculosis patients living in the Central Region of the Russian Federation. Resistance to RIF, INZ, and EMB was revealed by the absolute concentration test. The inhibitory concentration (IC) of EMB was determined for all the strains. Genetic changes in the above loci were estimated by mini-sequencing, followed by mass-spectrometry recording MALDI-TOF products. The relative low frequency of embB306 mutations was observed among the EMB-resistant strains (about 41.5%). Mutations in codon 306 were detected only in strains with EMB IC > or = 2 mg/L. A statistical significant association was found between the frequency of embB306 mutations and the multidrug resistant phenotype. A combination of these mutations with the traditional genetic markers of multidrug resistance may be used for the more effective detection of multidrug-resistant strains.

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[Vaccine prophylaxis of tuberculosis: implication and problems]. [Prevalence of tuberculosis among children and adolescents in the Russian Federation: analysis of official statistics]. [Analysis of the rates of infection and incidence of tuberculosis in children from a poor epidemic-situation area in 2001 to 2006]. [Clinical and epidemiological features of tuberculosis ostitis in BCG-vaccinated children]. [Detection of tuberculosis in Moscow adolescents].
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