摘要
目的探讨广东地区不同耐药表型的结核分枝杆菌临床分离株利福平耐药基因rpoB变异特征,为更精准快速分子检测技术的研发提供依据。方法从分枝杆菌菌种保藏库中筛选利福平(R)、异烟肼(H)、乙胺丁醇(E)、链霉素(s)敏感性明确的结核分枝杆菌临床分离株373株。对373株临床分离株采用聚合酶链式反应技术扩增包含利福平耐药决定区(RRDR)在内的rpoB基因片段并进行序列测定与分析。结果(1)373株菌株样本中,192株检测到rpoB基因变异,共发生225个突变包括碱基置换、插入、缺失在内的40种突变种类,531、526、511、516、533、515位点突变最为常见,占比分别为43.11%(97/225)、20.89%(47/225)、10.22%(23/225)、8%(18/225)、4.44%(10/225)、4%(9/225);(2)192例rpoB突变株中,159株仅发生单位点突变,33株发生双位点突变,其中D516V、H526L、H526D、H526R、S531W等14种突变种类只在单位点突变中出现,为单位点特有变异,M515T、H526Q、D516G、M515V、E541A等21种突变种类只在双位点突变中出现,为双位点特有变异,S531L、L511P、H526Y等5个种类突变为单双位点共有变异,不存在3个位点的同时变异。(3)对4个一线药物全敏感菌株(Q)、R敏感H耐药株(RSHR)、R耐药H敏感株(RRHS)和耐HR株(MDR)的rpoB基因突变率分别为4.67%(5/107)、28.42%(27/95)、90.62(29/32)、94.24%(131/139),除RRHS与MDR株间突变率不具统计学差异外(P>0.05),Q/R^(S)H^(R)、Q/R^(R)H^(S)、Q/MDR、R^(S)H^(R)/R^(R)H^(S)、R^(S)H^(R)/MDR间rpoB基因突变率具有统计学差异(均P<0.001)。结论广东地区结核分枝杆菌rpoB基因突变具有多样性、地域性特点,不同耐药表型可能对rpoB基因突变产生深远影响。
Objective To investigate the variation characteristics of rpoB gene of clinical isolates of Mycobacterium tuberculosis with different drug resistance phenotypes in Guangdong,so as to provide a research basis for the development of more accurate and rapid molecular detection technology.Methods 373 clinical isolates of Mycobacterium tuberculosis which had known their sensitivity to rifampicin(R),isoniazid(H),ethambutol(E)and streptomycin(S)were screened from the Mycobacterium collection.RpoB gene fragments including rifampin resistance determination region(RRDR)were amplified and sequenced from 373 clinical isolates by polymerase chain reaction.Results Firstly,among the 373 strains,192 strains were detected with rpoB gene variation,and a total of 225 mutations occurred in 40 types of mutation including base replacement,insertion and deletion.The mutations at 531,526,511,516,533 and 515 were the most common,accounting for 43.11%(97/225),20.89%(47/225),10.22%(23/225),8%(18/225),4.44%(10/225)and 4%(9/225)respectively.Secondly,among192 rpoB mutants,159 had only unit point mutation and 33 had double site mutation.Among them,14 mutation types such as D516V,H526L,H526D,H526R and S531W only appeared in unit point mutation,which was unit point specific variation.21 mutation types such as M515T,H526Q,D516G,M515V and E541A only appeared in double site mutation,which was double site specific.Five mutation types,including S531L,L511P,H526Y…etc.are single-and double-site common mutations.There is no simultaneous variation of three loci.Thirdly,the mutation rates of the rpoB gene for the four first-line drug fully susceptible strains(Q),R-sensitive and H-resistant strains(RSHR),R-resistant and H-susceptible strains(RRHS)and HR-resistant strains(MDR)were 4.67%(5/107),28.42%(27/95),90.62(29/32),94.24%(131/139),except that the mutation rate between RRHSand MDR strains was not statistically different(P>0.05),Q/R^(S)H^(R)、Q/R^(R)H^(S)、Q/MDR、R^(S)H^(R)/R^(R)H^(S)、R^(S)H^(R)/MDR strains were significantly different in rpoB gen
作者
孟繁荣
雷杰
牛群
王楠
吴玲
杨瑜
谢贝
李华
高俊文
傅红梅
刘志辉
MENG Fanrong;LEI Jie;NIU Qun;WANG Nan;WU Ling;YANG Yu;XIE Bei;LI Hua;GAO Junwen;FU Hongmei;LIU Zhihui(Institute of Pulmonary Diseases from Guangzhou Chest Hospital,Guangzhou 510095,China)
出处
《实用医学杂志》
CAS
北大核心
2022年第13期1582-1588,共7页
The Journal of Practical Medicine
基金
广东省自然科学基金项目(编号:2018A030313550)
广州市科技计划项目(编号:202002030152)
广东省医学科研基金项目(编号:A2020556)
广州市高水平临床重点专科和培育专科建设项目(编号:穗卫函[2019]1555号)
广州市医学重点学科(结核病学)建设项目。
关键词
结核分枝杆菌
RPOB基因
利福平耐药决定区
基因突变
Mycobacterium tuberculosis
rpoB gene
rifampicin resistance determination region
gene mutation
