对异烟肼与丙硫异烟胺耐药的结核分枝杆菌临床分离株检测及相关基因突变的研究  

A study on test and related genotypes of isoniazid-and-prothionamide-resistant Mycobacterium tuberculosis isolated from clinical specimens

作　　者：刘银萍 王杰 张俊仙 梁艳 李洪敏 吴雪琼 

机构地区：[1]解放军第三○九医院全军结核病研究所全军结核病防治重点实验室结核病诊疗新技术北京市重点实验室,北京100091

出　　处：《中国防痨杂志》2016年第9期718-721,共4页Chinese Journal of Antituberculosis

基　　金：基金项目：“十一五”国家重大传染病专项（No.2008ZX10003-001）;军队医学科技“十二五”重点项目（BWS11J050）;解放军第三.九医院课题（2014MS-012）

摘　　要：【摘要】目的研究MTB对异烟肼（INH）和丙硫异烟胺（Pto）的耐药情况，及从临床标本中的MTB临床分离株直接检测katG和inhA基因型的价值。方法回顾性调查解放军第三。九医院全军结核病研究所2014年8月至2015年8月住院确诊，并经抗结核药物治疗有效的结核病患者，共104例。患者临床标本经培养鉴定为MTB，然后通过绝对浓度法同时进行INH和Pto药物敏感性试验（简称“药敏试验”），并用基因芯片检测MTBkatG和inhA基因型。结果104例患者的MTB临床分离株经绝对浓度法药敏试验检测显示：20例（19．2％）对INH耐药，其中3例高度耐药、17例低度耐药；5例（4．8％）对Pro耐药，其中1例高度耐药、4例低度耐药；INH与Pto的交叉耐药率20．0％（4／20）。以传统药敏试验为对照，20例对INH耐药患者中，基因芯片检测10例（50．0％）发生katG基因315位点突变，3例（15．0%）发生inhA基因一15位点突变，1例（5．0％）发生双基因突变；84例INH敏感患者中，7例（8．3%）发生katG基因315位点突变，5例（6．0％）发生inhA基因-15位点突变。基因芯片检测katG基因315位点突变预示MTB对INH耐药的敏感度为55．0％（11／20），特异度为91．7％（77／84）；inhA基因-15位点突变预示MTB对INH和Pto耐药的敏感度分别为20．0%（4／20）和60．0％（3／5），特异度分别为94．0Y00（79／84）和93．9％（93／99）。结论大多数结核病患者的MTB临床分离株对INH和Pto耐药水平低，对Pto的耐药率低，katG315和inhA-15基因突变与MTB对INH耐药密切相关，intui-15基因突变与MTB对Pro耐药密切相关，应用基因芯片可快速检测临床标本中MTB的INH和Pto耐药基因型。Objective To investigate isoniazid （INH） and prothionamide （Pto） resistances in Mycobacterium tuberculosis （MTB）, and to assess the value of direct detection of katG and inhA genotypes in MTB isolated from clinical specimens. Methods A total of 104 hospitalized TB patients undergoing effective anti MTB treatment from Institute for Tuberculosis Research, the 309th Hospital of Chinese PLA between August 2014 and August 2015 were retrospectively studied. Clinical specimens were identified as MTB by culture media containing nitro- benzoic acid （PNB） and thiophene-carboxylic acid hydrazide （TCH） and INH and Pto drug susceptibility tests were performed at the same time by absolute concentration method. The mutations of MTI3 katG and inhA genes were detected using gene chip. Results Of the 104 clinical specimens, the results of drug susceptibility tests showed that 20 cases （19.20%） were INH resistant, in which 3 cases were with high-level resistant, and 17 cases were with low-level resistant; 5 cases （4.8%） were Pro-resistant, including 1 case with high-level and 4 cases with low level resistance. The cross-resistant rate between INH and Pto was 20.0% （4/20）. Compared to conventional drug susceptibility test, when using gene chip in the 20 TB patients with INH resistance, 10 cases （50.0%） had hatG315 mutations, 3 cases （15.0%） had inhA--15 mutations, and 1 case （5.00%o） had the mutations at hatG315 and inhA--15. Of the 84 INH sensitive TB patients, 7 cases （8.3%） had hatG315 mutations, 5 cases （6.0%） had i%tftA- 15 mutations. The sensitivity and specificity of hatG315 mutations detection by gene chip for 1NH resistance MTB were 55.0% （11/20） and 91.7% （77/84）, respectively. The sensitivities of inh4--15 mutation for INH and Pro resistance were 20.0% （4/20） and 60.0% （3/5）, respectively; and the specificities were 94.0% （79/84） and 93.9% （93/99）, respectively. Conclusion Most MTB isolated from clinical specimens were at low level of INH and Pro

关 键 词：分枝杆菌 结核 异烟肼 丙硫异烟胺 抗药性 细菌 芯片分析技术 

分 类 号：R378.11[医药卫生—病原生物学]