苏州市耐利福平结核分枝杆菌rpoB基因突变特点的研究  

Characterization of rpoB gene mutation in rifampin mycobacterium tuberculosis isolated from tuberculosis patients in Suzhou City

作　　者：张建平[1,2] 沈兴华[1,2] 王霞芳[1] 唐佩军[1,2] 虞忻[1,2] 曹丽娟[2] 王雪峰[2] 吴妹英[1] 

机构地区：[1]苏州大学附属传染病医院肺结核科,江苏苏州215007 [2]苏州大学基础医学与生物科学学院,江苏苏州215123

出　　处：《中华疾病控制杂志》2012年第4期293-296,共4页Chinese Journal of Disease Control & Prevention

基　　金：苏州市科技发展项目(SWKQ0824)

摘　　要：目的了解苏州市利福平(rifampin,RFP)耐药结核分枝杆菌rpoB基因突变特点,以期为苏州市RFP耐药结核分枝杆菌(mycobacterium tuberculosis,MTB)临床快速诊断提供有价值的分子标记靶点。方法通过裂解MTB抽提基因组DNA,采用PCR扩增和DNA测序方法,分析苏州市结核病患者痰培养阳性并经药敏试验诊断为RFP耐药MTB的rpoB基因RFP耐药决定区(rifampin resistance-determining region,RRDR)和CⅡ区变异谱与变异率。将主要突变形式RRDR-531(TCG→TTG)与传统药敏实验进行诊断学试验,评价此类型突变作为RFP耐药菌株分子标记靶点的临床应用价值。结果 42株RFP耐药MTB的rpoB基因总突变率为90.5%(38/42),在所检测到的突变中,以RRDR-531(TCG→TTG)为主要形式(61.9%)。以此类型的位点突变作为RFP耐药菌株诊断的分子标记,与传统药敏试验比较进行配对2检验,结果表明,与敏感菌株相比,RRDR-531(TCG→TTG)突变在RFP耐药菌株中差异有统计学意义(2=12.51,P<0.01)。结论 rpoB基因RRDR-531(TCG→TTG)突变是苏州市MTB产生RFP耐药性的主要分子机制,其作为RFP耐药MTB诊断的分子标记靶点具有较好的临床应用价值。Objective To understand the characteristics of rpoB gene mutation of rifampin（RFP） resistant Mycobacterium tuberculosis（MTB） isolated from tuberculosis patients in Suzhou City and the significance of potential typical mutation as a molecular marker to diagnose RFP-resistant MTB strains.Methods After dissolving MTB and amplifying rpoB gene fragment by PCR,direct sequencing was used to analyze the mutation spectrum occurred in rifampin resistance-determining region（RRDR） and CⅡ region of rpoB gene from 42 RFP-resistant and sensitive MTB strains.Diagnostic testing was used to evaluate the significance of typical mutation occurred in rpoB gene as a molecular marker of RFP-resistant MTB strains compared with the traditional sensitivity test.Results 38 of 42 RFP-resistant MTB strains（90.5%） was identified to contain base mutations occurred in RRDR an CⅡ region of rpoB gene.Among these,the mutation RRDR-531（TCG→TTG） was the dominating form and occupied 61.9%.When the mutation RRDR-531（TCG→TTG）was used to diagnose FRP-resistant MTB strains compared with the traditional sensitivity test as a marker,the mutation rate presented statistic significance（X2=12.51,P0.01）.Conclusions The mutation RRDR-531（TCG→TTG）was revealed to be the primary mechanism of MTB resistance to RFP and presented the significance in diagnosing RFP-resistant MTB strains as a molecular marker used in Suzhou City.

关 键 词：结核 分枝杆菌属 利福平 抗药性 

分 类 号：R378.911[医药卫生—病原生物学]