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

Abstract

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].Chinese journal of Epidemiology,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

Received:June 02, 2006

DOI：

KeyWord: 结核分枝杆菌耐药聚合酶链反应-单链构象多态性

English Key Word: Mycobacterium tuberculosis Drug resistance PCR-single-strand conformation polymorphism

FundProject:国家自然科学基金资助项目(30471526)

Author Name	Affiliation	E-mail
LIU Jing-hua	State Key Laboratory for Infectious Diseases Prevention and Control
ZHANG Li-shui	National Institute for Communicable Disease Control and Prevention
LIU Zhi-guang	State Key Laboratory for Infectious Diseases Prevention and Control
HUANG Ming-xiang	National Institute for Communicable Disease Control and Prevention
ZHAO Xiu-qin	State Key Laboratory for Infectious Diseases Prevention and Control
WANG Lin	National Institute for Communicable Disease Control and Prevention
WAN Kang-lin	State Key Laboratory for Infectious Diseases Prevention and Control	wankanglin@icdc.cn

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Abstract:

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。

English Abstract:

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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