基因芯片技术检测结核分枝杆菌对利福平和异烟肼耐药性的研究

目的探讨基因芯片技术在检测结核分枝杆菌利福平和异烟肼耐药性的应用。方法收集21例临床诊断结核病患者体液标本经培养得到的分离菌液,进行基因芯片检测,包括利福平的rpoB基因位点和异烟肼的KatG和inhA基因位点,从而判断其耐药性。结果 21例样本中,利福平rpoB基因有10例突变,总突变率为47.6%,其中9例为531位点的（C-T）突变,1例526位点的（C-G）突变。与药敏试验的符合率为81.0%。异烟肼基因突变有6例,总突变率为28.6%,其中4例为KatG 315（G-C）突变,2例为inhA-15（C-T）突变。与药敏试验的符合率为90.5%。利福平的11例耐药株中,基因芯片检测8例为突变型,突变率为72.7%,在利福平的10例敏感株中,基因芯片检测2例为突变型,突变率为20.0%。在异烟肼的6例耐药株中,基因芯片检测5例为突变型,突变率为83.3%,在异烟肼的15例敏感株中,基因芯片检测1例为突变型,突变率为6.7%。结论基因芯片技术检测结核分枝杆菌对利福平和异烟肼的耐药性具有较高的特异性和敏感性,快速准确,可以应用于结核分枝杆菌耐药性的检测。

The application of gene chip in detecting Mycobacterium tuberculosis resistance to rifampicin and isoniazid

Objective To investigate the application of gene chip in detecting Mycobacterium tuberculosis resistance to rifampicin(RFP) and isoniazid(INH).Methods Bacterium fluid cultured in the specimens of 21 patients with tuberculosis(TB) was examined for the resistance by gene chip,which included rpoB gene locus of rifampicin and KatG and inhA gene locus of isoniazid.Results 10 of 21 cases were found for the mutations of RFP rpoB gene,and the total mutation rate was 47.6%,which included 9 cases with 531(C-T) mutation and 1 case with 526(C-G) mutation.The coincidence rate of gene chip with medicine sensitivity tests was 81.0%.The 6 cases were found for the mutations of INH gene,and the total mutation rate was 28.6%,in which 4 cases with KatG 315(G-C) mutation and 2 cases with inhA-15(C-T) mutation.The coincidence rate of gene chip with medicine sensitivity tests was 90.5%.The mutation was tested by gene chip in 8 of 11 RFP resistance and the mutation rate was 72.7%.Mutation was found in 2 of 10 RFP susceptibility and the mutation rate was 20%.The 5 case of mutations were examined by gene chip in 6 cases INH resistance,and the mutation rate was 83.3%.1 case of mutation was observed in 15 cases of INH susceptibility and the mutation rate was 6.7%.Conclusion Gene chip in detecting Mycobacterium tuberculosis resistance to rifampicin and isoniazid has higher sensitivity and specificity,which is rapid and accurate and can be used to detect Mycobacterium tuberculosis resistance.