【摘要】 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。更多还原

【Abstract】 Objective To detect rifampicin(RFP) drug-resistant genes for mycobacterium tuberculosis by DNA micro-array chip and to provide experimental basis for clinical diagnosis and treatment. Methods A total of 1 005 sputumspecimens were collected to detect mycobacterium tuberculosis by direct smearing method from June 2013 to June 2015.Positive specimens were identified and isolated by modified Roche medium and DNA micro-array chip method, and drug-resistant genes were analyzed. Results The positive rate of mycobacterium tuberculosis was 40.3%(405/1 005) in 1 005 sputum specimens, and the positive rate 98.0%(397/405) detected by the DNA micro-array chip was higher than that detectedby the modified Roche culture(44.2%,179/405, P<0.01). The result of DNA micro-array chip showed that there were 90 strains mycobacterium tuberculosis with RFP drug-resistant, 33 strains(36.7%, 33/90) with simple rpo B gene mutation, 54strains(60.0%, 54/90) with rpo B and kat G gene mutation, and 3 strains(3.3%, 3/90) with rpo B and inh A gene mutation.Mutation sites of rpo B gene were in the order of 531, 526, 516, 511, and the mutation frequency was 44.4%(40/90), 25.6%(23/90), 10.0%(9/90), and 5.5%(5/90), respectively. The mutation types were 531(C->T), 531(C->G), 526(C->T), and 54.0%(29/90),and the frequency was 54.0%(29/90), 11.1%(10/90), 11.1%(10/90), respectively. Single point mutation and two pointmutation were 81 strains(90.0%, 81/90) and 9 strains(10.0%, 9/90). Conclusions The rpo B gene mutation is the molecularbasis for RFP drug-resistant of patients with pulmonary tuberculosis, and its mutations are diversity.更多还原