PCR-SSCP方法用于痰标本中结核分支杆菌耐药基因的检测

目的探讨采用直接检测结核病人痰标本中rpoB基因和KatG基因突变。评价该方法检测结核分支杆菌对利福平（RFP）和异烟肼（INH）耐药性。方法用PCR-SSCP技术分析，对35例耐INH（或含耐异烟肼）、63例耐RFP（或含耐RFP）的肺结核病人痰标本和20例非结核性肺部疾病组痰标本进行rpoB和KatG基因突变的检测。结果以结核分支杆菌H37RV为对照，63例耐RFP痰标本中57例PCR扩增阳性，其中46例SSCP图谱与H37Rv标准株有差异。rpoB突变率为65．2％。35例耐INH肺结核病人痰标本有20例扩增阳性。15例SSCP图谱与H37Rv标准株有差异，突变率为42．8％。20例非结核病人痰标本rpoB基因和KatG基因扩增均为阴性。结论PCR-SSCP技术有望成为结核分支杆菌耐药性检测的重要方法之一。

Detection of drug resistance gene of Mycobacterium tuberculosis in sputum specimens by PCR-SSCP

Objective To evaluate the value of detection of rpoB gene and katG gene mutation in Rifampin-resistant and Isoniadd-resistant Mycobactefium tuberculosis by PCR-SSCP.Methods 35sputum specimens from pulmonary tuberculous patients of Isoniazid-resistant,63sputum specimens from pulmonary tuberculous patients d Rifampin-resistant and 20 sputum specimeam from nontuberculous patients of pulmonary disease were detected by PCR-SSCP.Results SSCP pattern of Mycobactefium tuberculosis H37RV as conrtol .63sputum specinmns from pulmonary tubczcuhus of RFP-msistant were detected using PCR-SSCP,57 sputum specimens had positive PCR products,46 PCR products of sputum specimens had different SSCP patterns from the control. The rpoB mutation rate of sputum specimens was 65.2% using PCR-SSCP.35sputum specimens from pulmonary tuberculous of INH-resistant were detected using PCR-SSCP,20 sputum specimens had positive PCR products, 15 PCR products of sputum spcclnmns had different SSCP pattenns from the control. The katG mutation rote of sputum specimens was 42.8% .20sputum specimens from non-tuberculous patients of pulmonary disease,the rpoB gene and katG gene was negative. Conclusion PCR-SSCP may become a important method of detection the drag resistance of Mycobacterium tuberculosis.