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目的了解贵州地区结核分枝杆菌(MTB)对利福平耐药相关基因rpo B突变特征以便探讨适合本地区MTB对利福平耐药的快速筛选方法。方法用PCR-荧光探针法检测临床标本MTB核酸,阳性者(低浓度除外)用基因芯片检测rpo B基因,异常检测者再进行测序分析。结果 MTB核酸阳性患者3966例,阳性率22.72%(3966/17452)。基因芯片检测2738例中,552例有rpo B基因突变,突变率为20.16%(552/2738);9例异常检测者,经基因测序验证,分别为位点缺失、密码子插入、同密码子双碱基突变和多位点突变。rpo B基因突变类型有26种,以531,526,516,511,533,513和512位点突变为主,分别占56.23%,18.09%,9.67%,8.61%,4.57%,1.76%和0.18%。结论贵州地区MTB对利福平耐药相关基因rpo B突变特征与文献报道基本一致,基因芯片可作为本地区MTB利福平耐药的快速筛选方法。
Objective To understand the characteristics of rpo B gene mutation in rifampicin-resistant gene of Mycobacterium tuberculosis (MTB) in Guizhou Province in order to explore a rapid screening method for rifampin-resistant MTB in this region. Methods MTB nucleic acid was detected by PCR-fluorescence probe. The positive (except for low concentration) gene was detected by microarray. The rpo B gene was detected by DNA microarray. Results 3966 MTB nucleic acid positive patients, the positive rate of 22.72% (3966/17452). Among 2738 cases detected by gene chip, 552 cases had rpo B gene mutation with a mutation rate of 20.16% (552/2738). Nine cases were detected by gene sequencing, including site deletion, codon insertion, codon usage Double base mutation and multi site mutation. There were 26 types of rpo B gene mutation, with 531, 526, 516, 511, 533, 513 and 512 point mutations, accounting for 56.23%, 18.09%, 9.67%, 8.61%, 4.57%, 1.76% and 0.18% respectively. Conclusion The characteristics of rpo B gene mutation related to rifampicin resistance in Guizhou area are consistent with those reported in the literature. Gene chip can be used as a rapid screening method for rifampicin resistance in this area.