Molecular Characterization of Multidrug- and Extensively Drug-Resistant Mycobacterium tuberculosis Strains in Jiangxi, China
Molecular Characterization of Multidrug- and Extensively Drug-Resistant Mycobacterium tuberculosis Strains in Jiangxi, China
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DOI：
10.1128/jcm.06860-11
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发表时间：
2012-07-01
影响因子：
9.4
通讯作者：
Tu, Shaohua
中科院分区：
文献类型：
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作者：
Yuan, Xiaoliang;Zhang, Tiantuo;Tu, Shaohua
在本研究中，共有77株多重耐药和广泛耐药(分别为多药耐药和广谱耐药)的结核分枝杆菌分离自江西省中国。采用以下两种方法对所有耐药菌株进行基因分型：(1)采用分枝杆菌散布单位可变重复序列15位点法对所有耐药菌株进行基因分型，并用RD105缺失靶向多重聚合酶链式反应(RD105)鉴定北京家系；(2)用DNA测序法检测一线抗结核药物利福平(RIF)、异烟肼(INH)和二线药物氧氟沙星(OFX)、卡那霉素(Kan)、阿米卡星(AMK)和卷曲霉素(CAP)的耐药突变情况。检测了6个座位：RpoB(抗性)、katG和Maba-Inha(INH)、gyrA和gyrB(OFX)和RRs(Kan、AMK和CAP)。结果表明，这些菌株中以北京型为主(80.5%)，所选耐药株具有遗传多样性，可能具有独立获得性耐药性。与表型数据比较，DNA测序检测RIF、INH、OFX和Kan/AMK/CAP耐药性的敏感性分别为94.8%、80.5%、84.6%和78.9%。与RIF、INH、OFX和Kan/AMK/CAP耐药相关的突变分别为rpoB中Ser531Leu(44.2%)、katG中Ser315Thr(55.8%)和Maba-Inha中C-15T(11.7%)、gyra中Asp94Gly(48.7%)和RRS中A1401G(73.7%)。在rpoB基因中发现了5个新的katG突变体(Trp191Stop、Thr271Pro、Trp328Phe、Leu546Pro和Asp695Gly)和6个新等位基因(Ile569Val、Ile572Met、Phe584Ser、Val615Met、Asp626Glu和Lys972Thr)。
In this study, a total of 77 multidrug-resistant and extensively drug-resistant (MDR and XDR, respectively) isolates of Mycobacterium tuberculosis were characterized among samples from patients living in Jiangxi province, China. The following two approaches were used: (i) genotyping all drug-resistant isolates by the 15-locus MIRU-VNTR (mycobacterial interspersed repetitive- unit-variable-number tandem-repeat) method and identifying the Beijing family genotype using the RD105 deletion targeted multiplex PCR and (ii) determining the mutation profiles associated with the resistance to the first-line antituberculous drugs rifampin (RIF) and isoniazid (INH) and the second-line drugs ofloxacin (OFX), kanamycin (KAN), amikacin (AMK), and capreomycin (CAP) with DNA sequencing. Six loci were examined: rpoB (for resistance to RIF), katG and mabA-inhA (INH), gyrA and gyrB (OFX), and rrs (KAN, AMK, and CAP). It is shown that the Beijing genotype was predominant (80.5%) among these strains and that the selected drug-resistant strains were genetically diverse, suggesting that they probably had independently acquired drug resistance. In comparison to the phenotypic data, the sensitivities for the detection of RIF, INH, OFX, and KAN/AMK/CAP resistance by DNA sequencing were 94.8, 80.5, 84.6, and 78.9%, respectively. The most prevalent mutations involved in RIF, INH, OFX, and KAN/AMK/CAP resistance were Ser531Leu in rpoB (44.2%), Ser315Thr in katG (55.8%) and C-15T in mabA-inhA (11.7%), Asp94Gly in gyrA (48.7%), and A1401G in rrs (73.7%), respectively. Five novel katG mutants (Trp191Stop, Thr271Pro, Trp328Phe, Leu546Pro, and Asp695Gly) and six new alleles (Ile569Val, Ile572Met, Phe584Ser, Val615Met, Asp626Glu, and Lys972Thr) in the rpoB gene were identified.