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。
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.
出处
《中华流行病学杂志》
CAS
CSCD
北大核心
2006年第11期973-976,共4页
Chinese Journal of Epidemiology
基金
国家自然科学基金资助项目(30471526)
关键词
结核分枝杆菌
耐药
聚合酶链反应-单链构象多态性
Mycobacterium tuberculosis
Drug resistance
PCR-single-strand conformation polymorphism
作者简介
通讯作者：万康林，Email：wankanglin@icdc．cn
