用耐药基因检测方法指导老年肺结核病人治疗的临床研究

The clinical study on treatment of old patient with pulmonary tuberculosis by detecting drug resistance genes in Mycobacterium tuberculosis.

目的 了解老年肺结核病人的结核分支杆菌耐药情况 ,评价它们的临床应用价值。方法 采用聚合酶链反应 -单链构象多态性分析(PCR- SSCP)检测结核分支杆菌 rpo B、kat G、rps L、pnc A、emb B基因突变和药物敏感试验 (比例法 ) ,了解 1 1 7例老年肺结核病人结核分支杆菌耐药情况。探讨和比较 2种耐药性检测方法的检测效果。结果 1 /2以上的肺结核病人至少耐 2种抗结核药物 ,对 RFP、INH、SM、PZA和 EB总耐药率为82 .1 %、71 .7%、63.2 %、51 .2 %和 35.0 %。耐多药率为 67.5%。 rpo B、kat G、rps L、pnc A和 emb B基因突变率分别为 72 .8%、64.9%、59.8%、41 .0 %和 30 .7% ,多基因突变株达 76.0 %。结核杆菌耐药基因突变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区 ,也有少数基因突变发生在低耐药区。根据药敏试验和耐药基因检测 ,耐多药结核病人 6个月治愈率分别达到 54.8%和 62 .8% ,治疗效果满意 ,两种方法没有显著性差异 (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 ,可以快速检测结核杆菌 rpo B、kat G、rps L、pnc A和emb B耐药基因突变 ,可能会成为临床指导用药的好方法。

Objective To investigate drug resistance of Mycobacterium tuberculosis in old patients with pulmonary tuberculosis, To evaluate the clinical application value of detecting drug resistance genes in Mycobacterium tuberculosis. Methods Analyzing and comparing the results of mutation of genes and drug sensitive test in 117 M.tuberculosis clinical isolates by PCR SSCP detecting the rPOB?katG ?rPSL?PncA and embB resistance genes in M. tuberculosis and the proportion method on Lowenstein Jensen medium .Evaluating the clinical treatment effect.Results The rates of drug resistance to RFP? INH? SM ?PZA and EB were 82.1%?71.7%?63.2%?51.2%和35.0% respectively,. The gene mutation rate of RFP?INH?SM?PZA and EB were 72.8%?64.9%?59.8%?41.0%和30.7% respectively, The gene mutation was in relation with drug resistance level of M. Tuberculosis. The gene mutation rate was higher in high concentration drug resistance isolated strains than that in low concentration drug resistance isolated strains. The clinical cure rate of MDR TB 1n six months by drug sensitive test and by PCR SSCP was 54.8% and 62.8% ( P >0.05) , The treatment effect was satisfactory. Conclusions The method of detecting resistance genes of Mycobacterium tuberculosis was a new explore in guiding treatment, PCR SSCP is a sensitive and specific method for rapid detecting rpoB?katG?rpsL?pncA and embB genes mutations of M.TB. It might become good method for clinical management.