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

doi:10.3969/j.issn.1000-6621.2018.10.021

Abstract

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

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

Figures/Tables 3

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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