Preliminary study on clarithromycin resistance gene of Mycobacterium abscessus complex

doi:10.3969/j.issn.1000-6621.2021.12.013

Abstract

Abstract:

Objective To analyze the characteristics of clarithromycin resistance against Mycobacterium abscessus complex, and further to provide the basis for the adequate application and effective control of clarithromycin resistance in clinical. Methods A total of 385 Mycobacterium abscessus complex clinical isolates strains stored in National Tuberculosis Reference Laboratory of Chinese Center for Disease Control and Prevention for the national drug resistance surveillance from 2016 to 2017 were finally collected. Isolates were characterized using multilocus sequence analysis of various genes. Meanwhile, the drug susceptibility testing was determined using the broth microdilution method and the drug resistance genes were analyzed. Results Of all the 385 Mycobacterium abscessus complex isolates, 185 possessed the erm(41) T28 sequevar, 33 possessed the erm(41) C28 sequevar, 163 were M.massiliense, while the other four isolates belonged to M.bolletii. The resistance rate of amikacin against M.abscessus and M.massiliense was 2.8% (6/218) and 6.4% (14/218), respectively. For clarithromycin, the resistance rate was 4.3% (7/163) and 10.4% (17/163) for M.abscessus and M.massiliense, respectively.The resistance rate of clarithromycin against erm(41) C28 sequevar isolates was 9.1% (3/33) on day 3 and day 14 and no inducible resistance was occurred, however, the resistance rate was increased from 5.9% (11/185) on day 3 to 98.4% (182/185) on day 14 with 171 inducible resistant isolates of erm(41) T28 sequevar. Among the 31 acquired resistant isolates, 10 M.abscessus and 10 M.massiliense isolates possed an rrl 2058/2059 mutation with A2058C and A2058G the most common mutation type. Of the 9 amikacin resistant isolates with an A1408G rrs mutation, there were 7 isolates resistant to both clarithromycin and amikacin. Conclusion erm(41) T28 sequevar was more likely to induce clarithromycin resistance. The most common acquired resistance mutation site was A2058C for M.abscessus, which was A2058G in M.massiliense.

Key words: Mycobacteria,atypical, Genome,bacterial, Antimicrobial agent, Drug resistance, Drug evaluation

WANG Yi-ting, ZHENG Hui-wen, ZHAO Bing, XIA Hui, WANG Sheng-fen, OU Xi-chao, ZHOU Yang, SONG Yuan-yuan, ZHENG Yang, ZHAO Yan-lin, SHEN A-dong. Preliminary study on clarithromycin resistance gene of Mycobacterium abscessus complex[J]. Chinese Journal of Antituberculosis, 2021, 43(12): 1302-1307. doi: 10.3969/j.issn.1000-6621.2021.12.013

Figures/Tables 4

References 20

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引物名称	引物序列(5'~3')
16S rRNA-F	AGAGTTTGATCMTGGCTCAG
16S rRNA-R	CCGTCAATTCMTTTRAGTTT
rpoB-F	GGCAAGGTCACCCCGAAGGG
rpoB-R	AGCGGCTGCTGGGTGATCATC
hsp65-F	ATCGCCAAGGAGATCGAGCT
hsp65-R	AAGGTGCCGCGGATCTTGTT
转录间隔区-F	AAGTCGTAACAAGGTARCCG
转录间隔区-R	TCGCCAAGGCATCCACC
erm(41)-F	GACCGGGGCCTTCTTCGTGAT
erm(41)-R	GACTTCCCCGCACCGATTCC
rrl-F	GTAGCGAAATTCCTTGTCGG
rrl-R	TTCCCGCTTAGATGCTTTCAG
rrs-F	ATGACGTCAAGTCATCATGCC
rrs-R	AGGTGATCCAGCCGCACCTTC

药品	MIC范围(μg/ml)			耐药菌株[株(耐药率,%)]
药品	敏感	中度	耐药	脓肿分枝杆菌(218株)	马赛分枝杆菌(163株)
阿米卡星	≤16	32	≥64	6(2.8)	7(4.3)
克拉霉素	≤2	4	≥8	14(6.4)	17(10.4)
阿奇霉素	≤16	-	≥32	33(15.1)	21(12.9)
利奈唑胺	≤8	16	≥32	34(15.6)	36(22.1)
替加环素	≤1	2~4	≥8	50(22.9)	56(34.3)
头孢西丁	≤16	32~64	≥128	56(25.7)	51(31.3)
加替沙星	≤1	2	≥4	99(45.4)	102(62.6)
莫西沙星	≤1	2	≥4	120(55.0)	108(66.3)
妥布霉素	≤2	4	≥8	135(61.9)	148(90.8)
亚胺培南	≤4	8~16	≥32	181(83.0)	126(77.3)
米诺环素	≤1	2~4	≥8	198(90.8)	83(50.9)
利福布汀	≤0.5	-	≥1	212(97.2)	154(94.5)
美罗培南	≤4	8~16	≥32	213(97.7)	159(97.5)
左氧氟沙星	≤1	2	≥4	216(99.1)	155(95.1)

菌株	rrl 基因突变(株)					rrs基因A1408G 突变(株)
菌株	A2058C	A2058G	A2059C	合计	突变率(%)	rrs基因A1408G 突变(株)
脓肿分枝杆菌
erm(41)T28型	4	2	1	7	22.6	3
erm(41)T28型	-	-	-	-	-	2
erm(41)C28型	2	1	0	3	9.7	2
马赛分枝杆菌	3	5	2	10	32.3	2