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目的用DNA微阵列芯片法检测结核分枝杆菌耐药基因,为临床诊治提供实验依据。方法收集2013年6月—2015年6月1 005份痰标本,采用直接涂片检测结核分枝杆菌情况,阳性样本分别用改良罗氏培养基培养和DNA微阵列芯片法进行鉴定,分析耐药基因情况。结果 1 005份痰标本的涂阳率为40.3%(405/1 005)。DNA微阵列芯片法的阳性率为98.0%(397/405),明显高于改良罗氏培养基培养法(44.2%,179/405),P<0.01。DNA微阵列芯片法结果显示,有90株(24.9%,90/361)结核分枝杆菌发生利福平耐药,其中33株(占36.7%)发生单纯rpo B基因突变,54株(占60.0%)发生rpo B和kat G基因突变,3株(占3.3%)发生rpo B和inh A基因突变;rpo B基因最常见的突变位点依次为531、526、516、511,突变频率分别为44.4%、25.6%、10.0%、5.5%;突变频率最高的三种突变类型依次为:531位点(C->T)占54.0%(29/90),531位点(C->G)占11.1%(10/90)及526位点(C->T)占11.1%(10/90);单位点突变和二位点联合突变分别有81株(占90.0%)和9株(占10.0%)。结论 rpo B基因突变是导致肺结核患者RFP耐药的分子基础,其突变位点呈多样性。
Objective To detect the drug resistance genes of Mycobacterium tuberculosis by DNA microarray and provide experimental evidence for clinical diagnosis and treatment. Methods A total of 1 005 sputum samples were collected from June 2013 to June 2015. Mycobacterium tuberculosis was detected by direct smear. The positive samples were identified by modified Roche medium and DNA microarray. Happening. Results The smear rate of 1 005 sputum samples was 40.3% (405/1 005). The positive rate of DNA microarray was 98.0% (397/405), which was significantly higher than that of modified Roche culture (44.2%, 179/405) (P <0.01). DNA microarray chip method showed that there were 90 (24.9%, 90/361) Mycobacterium tuberculosis rifampicin-resistant, of which 33 (36.7%) occurred a simple rpo B gene mutation, 54 strains (60.0 %) Occurred rpo B and kat G gene mutations, 3 (3.3%) occurred rpo B and inh A gene mutations; rpo B gene the most common mutation sites were 531,526,516,511, the mutation frequency was 44.4%, 25.6%, 10.0% and 5.5%, respectively. The three mutation types with the highest mutation frequency were 54.0% (29/90) and 531 (C-> G) Accounting for 11.1% (10/90) and 52.1 (C> T), accounting for 11.1% (10/90). There were 81 strains (90.0%) and 9 strains 10.0%). Conclusion The mutation of rpo B gene is the molecular basis of RFP resistance in patients with pulmonary tuberculosis. The mutations of rpo B gene are diverse.