结核分枝杆菌利福平耐药基因rpoB突变特征初步分析

刘敬华; 张丽水; 刘志广; 黄明翔; 赵秀芹; 王琳; 万康林

文章摘要

刘敬华,张丽水,刘志广,黄明翔,赵秀芹,王琳,万康林.结核分枝杆菌利福平耐药基因rpoB突变特征初步分析[J].中华流行病学杂志,2006,27(11):973-976

结核分枝杆菌利福平耐药基因rpoB突变特征初步分析

Study on the characteristics of mutation on Mycobacterium tuberculosis rifampicin-resistance gene

收稿日期:2006-06-02 出版日期:2014-09-12

DOI：

中文关键词: 结核分枝杆菌耐药聚合酶链反应-单链构象多态性

英文关键词: Mycobacterium tuberculosis Drug resistance PCR-single-strand conformation polymorphism

基金项目:国家自然科学基金资助项目(30471526)

作者	单位	E-mail
刘敬华	中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室 102206
张丽水	福建省福州市肺科医院
刘志广	中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室 102206
黄明翔	福建省福州市肺科医院
赵秀芹	中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室 102206
王琳	福建省福州市肺科医院
万康林	中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室 102206	wankanglin@icdc.cn

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中文摘要:

目的阐明结核分枝杆菌利福平耐药株rpoB基因突变特点。方法对110株结核分枝杆菌临床分离株包括rpoB基因利福平耐药决定区(RRDR)81个碱基在内的286 bp片段进行聚合酶链反应-单链构象多态性(PCR-SSCP)分析,并对经检测显示有突变的结核分枝杆菌rpoB基因此扩增区域进行序列测定。其中利福平耐药株73株,非利福平耐药株11株,敏感株26株。结果PCR-SSCP检测显示利福平耐药株中有47株有突变存在,经直接测序分析其中76．6%的菌株存在rpoB基因的单位点突变,主要集中在531位(61．1%)和526位(25．0%)；联合突变发生率为23．4%。此外,经PCR-SSCP检测,11株非利福平耐药株中2株有突变存在,26株敏感株中1株有突变存在。经直接测序分析突变涉及位点为511位、526位和535位。结论rpoB基因耐药决定区发生突变是结核分枝杆菌利福平耐药的主要原因,其中531位丝氨酸和526位组氨酸是最常见的突变位点。

英文摘要:

Objective To elucidate the characters of rpoB mutation in rifampin-resistant clinical isolates of Mycobacterium tuberculosis.Methods 286 bp DNA fragment of rpoB gene including 81 bp code region (rifampin resistance deteremination region,RRDR)was analyzed by PCR-single-strand conformation polymorphism(SSCP).The 286 bp DNA fragment of each strain which had been proved to have mutation by PCR-SSCP was then sequenced.110 strains of M.tuberculosis,including 73 rifampin- resistant strains,11 rifampin-susceptible drug-resistant strains and 26 drug-susceptible strains were studied. Results 47 rifampin-resistant strains were detected to have mutations by PCR-SSCP method.76.6% rifampin-resistant strains showed that rpoB gene was carrying single point mutation analyzed by direct sequencing technique,which mainly located at 531-Ser(61.1%) and 526-His (25.0%).The combined mutation rate was 23.4 %.In addition,2 rifampin-susceptible drug-resistant strains and 1 drug-susceptible strain were mutated,detected by PCR-SSCP method.Sequencing results showed that the mutations located at 511-Leu,526-His and 535-Pro.Conclusion Mutations in the 81 bp RRDR of rpoB were the main reasons of M.tuberculosis resistant to rifampin.531-Ser and 526-His were the most common positions of mutations

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