两种快速方法联合检测结核分枝杆菌耐药性

目的建立联合检测结核分枝杆菌耐药性试验方法,以提高耐药检测方法的灵敏度和特异度。方法对126株已知耐药浓度的结核分枝杆菌耐药株和敏感株分别采用硝酸盐还原试验(Nitrate Reductase Assay,NRA)和反向斑点杂交试验(reverse-dot blot hybridization,RBH),检测其对异烟肼(INH)、利福平(RFP)、链霉素(SM)和乙胺丁醇(EMB)的耐药性,并将结果与绝对浓度法结果相比较。结果硝酸盐还原试验和反向斑点杂交试验联合检测与绝对浓度法比较具有高度的一致性(P﹥0.05),异烟肼灵敏度95.5%,特异度100%;利福平灵敏度100%,特异度98.1%;链霉素灵敏度93.2%,特异度92.7%;乙胺丁醇灵敏度94.1%,特异度94.6%。结论硝酸盐还原试验和反向斑点杂交试验联合检测可作为快速结核分枝杆菌耐药性试验方法。

Joint detection on drug resistance of mycobacterium tuberculosis with Nitrate Reductase Assay and Reverse-dot Blot Hybridization

OBJECTIVE To establish a joint detection method for drug resistance of mycobacterium tuberculosis,to improve the sensitivity and specificity of resistance detection methods.METHODS 126 resistant and sensitive strains of MTB were analyzed by Nitrate Reductase Assay(NRA)and Reverse-dot Blot Hybridization(RBH)for isoniazid(INH)and rifampicin(RFP)respectively.The results were compared with absolute drug susceptibility tests.RESULTS The joint detection had a relatively high degree of consistency(P﹥0.05),compared Nitrate Reductase Assay,Reverse-dot Blot Hybridization and the joint detection with absolute concentration method.The sensitivity of INH,RPF,SM and EMB was 95.5%,100%,93.2% and 94.1%,and the specificity was 100%,98.1%,92.7% and 94.6% respectively.CONCLUSION The Nitrate Reductase Assay and Reverse-dot Blot Hybridization can be used as fast joint detection on drug resistance of mycobacterium tuberculosis.