结核分支杆菌五种耐药基因检测的临床应用及评价

目的　检测结核分支杆菌rpoBkatG、rpsL、pncA和embB耐药基因 ,评价其临床应用价值。方法　采用聚合酶链反应 单链构象多态性 (PCR SSCP)分析和药敏试验 (比例法 ) ,了解 10 9例肺结核患者结核分支杆菌耐药情况 ,并分析、比较临床治疗效果。结果　 1/ 2以上的肺结核患者至少耐两种抗结核药物 ,对RFP、INH、SM、PZA和EB总耐药率分别为 80 7%、71 5 %、78 8%、5 7 7%和48 6%。rpoB、katG、rpsL、pncA和embB基因突变率分别为 76%、68%、71%、5 1%和 3 0 %。结核分支杆菌耐药基因突变率与耐药水平联系密切 ,多数结核分支杆菌耐药基因突变易发生在高耐药株 ,也有少数基因突变发生在低耐药株。根据药敏试验和耐药基因检测结果 ,6个月耐多药结核治愈率分别达到 5 4 8%和 62 8% ,治疗效果满意 ,两种方法差异无显著性 (P >0 0 5 )。结论　耐药基因检测指导治疗是一种新探索 ,PCR SSCP方法敏感、特异 ,可以快速检测结核分支杆菌rpoB、katG、rpsL、pncA和embB耐药基因突变 ,可能会成为临床指导用药的好方法

Clinical application and evaluation of the detection of five drugs resistance genes in Mycobacterium tuberculosis

Objective To evaluate the clinical application of the detection of the drug resistant genes rPOB, katG, rPSL, PncA and embB in M tuberculosis by using polymerase chain reaction single strand conformation polymorphism (PCR SSCP) analysis Methods Gene mutations and anti tuberculous drug susceptibility test were analyzed in 109 M tuberculosis isolates by PCR SSCP and the proportion method on Lowenstein Jensen medium, respectively The therapeutic effect was evaluated in patients with tuberculosis Results Isolates from more than 50% of the patients with pulmonary tuberculosis were resistant to at least two drugs The total rates of drug resistance to rifampin (RFP), isoniazid( INH), streptomycin ( SM), pyrizinamide ( PZA ) and ethambutol (EB) were 80 7%, 71 5%, 78 8%, 57 7%, and 48 6%, respectively The gene mutation rates of rpoB,katG,rpsL,pncA and embB were 76%,68%,71%,51% and 30%,respectively The gene mutations were correlated with the degree of drug resistance to M tuberculosis Most of the gene mutations were found in drug resistant isolates in high concentrations The six month cure rates of MDR TB, confirmed by drug susceptibility test and by PCR SSCP,were 54 8% and 62 8%, respectively ( P >0 05) Conclusions PCR SSCP is a sensitive and specific method for rapid detection of rpoB,katG,rpsL,pncA and embB gene mutations in M tuberculosis Drug resistant gene detection may be clinically useful in the therapy of tuberculosis