MTB对阿米卡星和卡那霉素耐药相关rrs与eis基因突变的研究

doi:10.3969/j.issn.1000-6621.2018.10.021

中国防痨杂志 ›› 2018, Vol. 40 ›› Issue (10): 1141-1143.doi: 10.3969/j.issn.1000-6621.2018.10.021

MTB对阿米卡星和卡那霉素耐药相关rrs与eis基因突变的研究

李可维¹,赵薇^1,^†(),刘思洁^1,²,杨修军¹,申莉³,刘昕²,齐鹏²

^1. 130062 长春,吉林省疾病预防控制中心微生物检验所
^2. 吉林省结核病防治科学研究院结核实验室
^3. 吉林大学公共卫生学院流行病与卫生统计学教研室

收稿日期:2018-09-11 出版日期:2018-10-10 发布日期:2018-10-18
通信作者: 赵薇 E-mail:weizhao81226@126.com
基金资助:
吉林省卫生和计划生育委员会技术创新项目(2016J036)

Study on mutations of rrs and eis genes in Mycobacterium tuberculosis resistant to amikacin and kanamycin

Ke-wei LI¹,Wei ZHAO^1,^†(),Si-jie LIU^1,²,Xiu-jun YANG¹,Li SHEN³,Xin LIU²,Peng QI²

^1. Microbiological Laboratory, Jilin Center for Disease Control and Prevention, Changchun 130062,China

Received:2018-09-11 Online:2018-10-10 Published:2018-10-18
Contact: Wei ZHAO E-mail:weizhao81226@126.com

摘要/Abstract

摘要：

本研究搜集了吉林省全部9个地级市结核病医疗机构2014—2017年内1357株结核分枝杆菌分离株。采用比例法进行Am和Km的药物敏感性试验,并对rrs和eis基因进行PCR和测序分析。发现28株同时耐Km和Am,单耐Km者有2株; 其中26株MTB的rrs基因均发生突变,基因突变率为92.86%;6株菌存在eis基因突变,突变率为21.43%。60株对照株未发现基因突变。吉林省MTB对Am和Km存在较高的交叉耐药现象;rrs基因的突变可以作为两者交叉耐药的分子标志,为建立耐Am和(或)Km结核病快速检测技术提供分子基础。

关键词: 分枝杆菌, 结核, 多药耐药相关蛋白质类, 丁胺卡那霉素, 卡那霉素, DNA突变分析, rrs基因, eis基因

Abstract:

1357 Mycobacterium tuberculosis (MTB) isolates were collected from tuberculosis medical institutions in nine prefectures of Jilin Province from 2014 to 2017. Amikacin (Am) and kanamycin (Km) susceptibility tests were performed by proportional method, and rrs and eis genes were analyzed by PCR and sequencing. Twenty-eight strains were resistant to both Km and Am, and 2 strains were resistant to Km alone. rrs gene mutation occurred in 26 strains resistant to Am and Km, and the mutation rate was 92.86%. eis gene mutation occurred in 6 strains, and the mutation rate was 21.43%. No mutation was found in 60 control strains. MTB had high cross resistance to Am and Km in Jilin Province. The mutation of rrs gene can be used as a molecular marker of cross resistance, which can provide molecular basis for the establishment of rapid detection technology for tuberculosis resistant to Am and/or Km.

Key words: Mycobacterium tuberculosis, Multidrug resistance-associated proteins, Amikacin, Kanamycin, DNA mutational analysis, rrs gene, eis gene

李可维,赵薇,刘思洁,杨修军,申莉,刘昕,齐鹏. MTB对阿米卡星和卡那霉素耐药相关rrs与eis基因突变的研究[J]. 中国防痨杂志, 2018, 40(10): 1141-1143. doi: 10.3969/j.issn.1000-6621.2018.10.021

Ke-wei LI,Wei ZHAO,Si-jie LIU,Xiu-jun YANG,Li SHEN,Xin LIU,Peng QI. Study on mutations of rrs and eis genes in Mycobacterium tuberculosis resistant to amikacin and kanamycin[J]. Chinese Journal of Antituberculosis, 2018, 40(10): 1141-1143. doi: 10.3969/j.issn.1000-6621.2018.10.021

图/表 3

参考文献 18

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基因	上下游引物名称	引物序列(5'~3')	退火温度(℃)	产物大小(bp)
rrs	F	GTCCGAGTGTTGCCTCAGG	60	516
	R	GTCAACTCGGAGGAAGGTGG
eis	F	GCGTAACGTCACGGCGAAATTC	60	567
	R	GTCAGCTCATGCAAGGTG

突变位点	突变数(株)	突变率(%)
57	2	7.14
114	2	7.14
379	2	7.14
505	2	7.14

突变类型	碱基改变	A/A 改变	突变数(株)
单位点碱基突变
57	CCT→CCA	无	2
114	CAG→CAA	无	2
双位点碱基突变
379,505	GCG→CCG, CGG→GGG	A→P, R→G	2

MTB对阿米卡星和卡那霉素耐药相关rrs与eis基因突变的研究

Study on mutations of rrs and eis genes in Mycobacterium tuberculosis resistant to amikacin and kanamycin

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