High prevalence of KatG Ser315Thr substitution among isoniazid-resistant Mycobacterium tuberculosis clinical isolates from northwestern Russia, 1996 to 2001
High prevalence of KatG Ser315Thr substitution among isoniazid-resistant Mycobacterium tuberculosis clinical isolates from northwestern Russia, 1996 to 2001
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DOI：
10.1128/aac.46.5.1417-1424.2002
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发表时间：
2002-05-01
影响因子：
4.9
通讯作者：
Vyshnevskiy, B
中科院分区：
文献类型：
--
作者：
Mokrousov, I;Narvskaya, O;Vyshnevskiy, B
共204异烟肼（INH）耐药结核分枝杆菌菌株分离自不同的患者在俄罗斯西北部地区从1996年至2001年进行了筛选的PCR-限制性片段长度多态性（RFLP）分析。该测定使用katG基因的扩增片段的HapII切割来检测颠换315AGC-> ACC（Ser-> Thr），其与INH抗性相关。该分析显示，来自新诊断和既往诊断的结核病（TB）患者的菌株中katG S315T突变的患病率为93.6%。在研究中纳入的57株INH敏感分离株中均未发现该突变。该检测试剂盒的特异性为100%;所有含有S315T突变的分离株均通过基于培养物的药敏试验方法归类为耐药。IS6110-RFLP分析和间隔区寡核苷酸分型（spoligotyping）方法确定的北京基因型，发现在60.3%的INH耐药株研究。katG S315 T移位在北京基因型菌株中比在非北京基因型菌株中更普遍：1999年至2001年分离的所有分离株（包括来自新发病例和既往诊断病例的分离株）分别为97.8%和84.6%，1996年至2001年分离的来自新发病例的分离株分别为100.0%和86.5%。该PCR-RFLP分析的设计允许快速和明确地鉴定katG 315ACC突变等位基因。该检测方法的简单性允许其在结核病高发地区（包括俄罗斯西北部）的临床微生物学实验室的常规实践中实施，其中该突变占主导地位。
A total of 204 isoniazid (INH)-resistant strains of Mycobacterium tuberculosis isolated from different patients in the northwestern region of Russia from 1996 to 2001 were screened by a PCR-restriction fragment length polymorphism (RFLP) assay. This assay uses HapII cleavage of an amplified fragment of the katG gene to detect the transversion 315AGC-->ACC (Ser-->Thr), which is associated with INH resistance. This analysis revealed a 93.6% prevalence of the katG S315T mutation in strains from patients with both newly and previously diagnosed cases of tuberculosis (TB). This mutation was not found in any of 57 INH-susceptible isolates included in the study. The specificity of the assay was 100%; all isolates that contained the S315T mutation were classified as resistant by a culture-based susceptibility testing method. The Beijing genotype, defined by IS6110-RFLP analysis and the spacer oligonucleotide typing (spoligotyping) method, was found in 60.3% of the INH-resistant strains studied. The katG S315T shift was more prevalent among Beijing genotype strains than among non-Beijing genotype: 97.8 versus 84.6%, respectively, for all isolates, including those from patients with new and previously diagnosed cases, isolated from 1999 to 2001 and 100.0 versus 86.5%, respectively, for isolates from patients with new cases isolated from 1996 to 2001. The design of this PCR-RFLP assay allows the rapid and unambiguous identification of the katG 315ACC mutant allele. The simplicity of the assay permits its implementation into routine practice in clinical microbiology laboratories in regions with a high incidence of TB where this mutation is predominant, including northwestern Russia.