运用反向斑点杂交技术检测四川地区结核分枝杆菌katG基因315位点的突变

为了检测四川地区结核分枝杆菌KatG基因315位点的突变特点,并建立耐异烟肼结核分枝杆菌快速检测方法。运用反向杂交技术检测四川地区65株耐异烟肼和10株异烟肼敏感结核分枝杆菌KatG基因315位点的突变,并用DNA直接测序法验证杂交结果。结果显示在65株耐异烟肼结核分枝杆菌中,发现41株在KatG基因315位点发生突变,突变率最大的是AGC→ACC,占90％,其次是AGC→AAC和AGC→ATC突变。DNA序列测定结果进一步验证了反向杂交的结果。运用反向杂交技术检测四川地区结核分枝杆菌KatG基因315位点的突变的检出率和特异性分别是63％和100％。反向斑点杂交技术能快速有效地检测四川地区结核分枝杆菌KatG基因315位点的突变。

Detection of Mutations at Codon 315 of KatG gene in Isoniazid-resistant Mycobacterium tuberculosis Isolates from Sichuan Region by Reverse Blot Hybridization

Reverse blot hybridization (RBH) was applied to examine S315 mutations within katG gene in 65 INH resistant and 10 INH susceptible M. tuberculosis isolates. DNA sequencing was utilized to validate the RBH results. The Results showed that mutations at codon 315 in katG gene were detected in 41 of the 65 (63%) INH resistant M. tuberculosis isolates; the most frequent mutation type was AGC→ACC occurring in 37 (90%) isolates, followed by AGC→AAC and AGC→ATC. The results of DNA sequencing further confirmed the RBH results. The sensitivity and specificity of RBH array by S315 mutation testing were 63% and 100%, respectively. The S315 mutation in katG gene in M. tuberculosis isolates recovered from Sichuan region showed geographical variation.