结核分支杆菌rpoB基因套式PCR在利福平耐药和结核病辅助诊断中的应用

目的 分析耐利福平的结核分支杆菌(Mtb)rpoB基因突变状况。方法 对38株耐利福平Mtb菌株的rpoB基因突变高频区域进行套式PCR扩增,并进一步全自动测序。结果 38株耐利福平Mtb中有36株存在突变(包含5个密码子11种形式的突变),突变率94．7％。结论 Mtb xpoB基因的突变与利福平耐药高度关联,rpoB基因套式PCR及测序能快速、正确鉴别耐利福平菌株。

Application of rpoB gene nested polymerase chain reaction in Mycobacterium tuberculosis strains resistant to rifampin and assistant diagnosis for the patients with tuberculosis

Objective To analyze rpoB mutations of Mycobacterium tuberculosis strains with rifampin resistance isolated from the patients with tuberculosis. Methods By nested polymerase chain reaction(nPCR),the fragment including a high frequency mutation region of ropB was amplified from 38 M.tuberculosis strains resistant to rifampin ,the fragments were sequenced by automated sequencing machine. Results Eleven types of mutations involving 5 codons in the rpoB gene were identified in 36 rifampin resistant strains(94.7%). Conclusion There is a strong correlation between resistance to rifampin and rpoB mutations of M.tuberculosis, the sequencing analysis of the rpoB fragment amplified by nPCR may rapidly and accurately identify the rifampin resistant strains of M. tuberculosis.