线性探针杂交技术在广西百色地区耐药结核病检测中的应用

目的 了解广西百色地区利福平、异烟肼耐药基因的分布情况。评价线性探针杂交技术(LPA)检测耐药结核分枝杆菌的应用价值。方法 利用LPA技术与传统比例药敏试验分别检测496例结核分枝杆菌感染患者痰标本,统计出利福平、异烟肼各个耐药基因的分布情况并与传统比例药敏试验作比较。结果 LPA技术对结核病患者耐多药的检出率和传统比例药敏试验相比较差异无统计学意义(P>0.05) 。LPA技术检测耐利福平和异烟肼的灵敏度、特异度分别为97.0%、99.1%和77.8%、 99.5%,耐多药结核病的灵敏度和特异度分别为82.8%和99.8%。本地区利福平耐药基因突变频率最高的是rpoB WT8 占32.4%,其次是ropB MUT3占29.7%;异烟肼耐药基因最常见的突变是 katG WT 占46.8%,其次是katG MUT1 占43.6%。kappa一致性检验评价两种药物耐药性检测方法的kappa值分别为0.965和0.904。结论 广西百色地区结核分枝杆菌利福平的耐药基因以rpoB WT8和ropB MUT3两个位点突变为主,而结核分枝杆菌异烟肼的耐药基因则以katG WT和katG MUT1位点突变多见。LPA技术能快速、准确诊断结核分枝杆菌利福平、异烟肼的耐药性及多药耐药性,LPA技术检测结核分枝杆菌利福平和异烟肼耐药性与传统比例药敏试验具有高度的一致性,LPA检测技术可以大大地缩短检测的时限,辅助临床快速诊断出多耐药的结核菌。

Application of linear probe hybridization method in analysis of multi-drug resistant TB in Baise area,Guangxi

Objective To study the distribution of resistant genes about RFP and INH in patients with tuberculosis in Baise area, Guangxi Province, and evaluate the application of molecular linear probe hybridization method ( LPA) in rapid detection of multi-drug resistant tuberculosis. Methods The LPA method was used to detect 496 cases of Mycobacterium tuberculosis clinical isolates. At the same time, the traditional method was also used to detect and compare these samples. Results The differences of multi-drug resistance detection rate among LPA method and traditional method were not statistically significant (P>0.05). The sensitivity and specificity of RFP and INH clinical isolates were 97.0%, 99.1% and 77.8%, 99.5% respectively by LPA method. The sensitivity and specificity of MDR by LPA method were 82.8% and 99.8% respectively, and the mutant resistant strains of RFP mainly distributed in rpoB WT8 32.4%, ropB MUT3 29.7%, and INH were katG WT 46.8% and katG MUT1 43.6%. The values of kappa were 0.965 and 0.904 for the two methods. Conclusion RpoB WT8 and ropB MUT3 can be mostly seen in the mutant resistant strains of RFP, and as well as katG WT and katG MUT1 in the mutant resistant strains of INH. LPA method can detect RFP and INH drug resistance and other common resistant genes rapidly and accurately, which means it can sharply reduce the susceptibility test time for timely clinical drug treatment.