Subspecies and in vitro drug sensitivity analysis of Mycobacterium intracellular clinical isolates

doi:10.3969/j.issn.1000-6621.2021.02.009

Abstract

Abstract:

Objective To analyze the subspecies composition and drug resistance profiles of Mycobacterium intracellular clinical isolates in Shenzhen, providing scientific basis for the treatment of Mycobacterium intracellular pulmonary disease. Methods A total of 97 Mycobacterium intracellular clinical strains stored in National Tuberculosis Reference Laboratory of Chinese Center for Disease Control and Prevention were finally included. All strains were isolated from respiratory specimens of suspected tuberculosis or NTM patients in Shenzhen Third People’s Hospital in 2018. Multi-target sequencing was applied to subspecies identification and minimum inhibitory concentrations (MIC) of strains to 13 drugs (clarithromycin, amikacin, moxifloxacin, linezolid, rifampin, rifabutin, ethambutol, streptomycin, doxycycline, ciprofloxacin, isoniazid, ethionamide and trimethoprim/sulfamethoxazole) was tested by Sensititre^? SLOWMYCO plate. Results The majority subspecies of Mycobacterium intracellular was Mycobacterium intracellular subsp,accounting for 63.92% (62/97), followed by Mycobacterium intracellular subsp.chimaera (18.56% (18/97)) and Mycobacterium intracellular subsp.paraintracellulare (17.53% (17/97)). Drug susceptibility testing (DST) indicated that the lowest values of MIC₅₀ and MIC₉₀ were 0.5 and 8 μg/ml (rifabutin); and the highest were 32 and 64 μg/ml (streptomycin) in Mycobacterium intracellular. The value of MIC₉₀ of clarithromycin in Mycobacterium intracellular and Mycobacterium paraintracellular both was 2 μg/ml while that in Mycobacterium chimaera was 64 μg/ml. The drug resistant rates of Mycobacterium intracellular to clarithromycin, ethambutol, rifampicin, rifabutin, streptomycin, amikacin, linezolid and moxifloxacin were 9.28% (9/97), 44.33% (43/97), 42.27% (41/97), 15.46% (15/97), 36.08% (35/97), 11.34% (11/97), 38.14% (37/97) and 46.39% (45/97), respectively. The resistance of Mycobacterium chimaera to clarithromycin was significantly higher than that of Mycobacterium paraintracellulare (27.78% (5/18) vs. 4.84% (3/62), χ²=8.156, P=0.012). Conclusion The dominant subspecies was Mycobacterium intracellular subsp. in Mycobacterium intracellular. Drug resistance profiles varied a lot within subspecies to different drugs, thus subspecies identification and DST should be conducted to guide tailored therapy.

Key words: Mycobacteria,atypical, Mycobacterium chelonae, Bacterial typing techniques, Microbial sensitivity tests, Drug resistance,bacterial

LI Yuan-chun, ZHANG Yue, ZENG Xiang-jie, HE Wen-cong, QIU Qian, ZHAO Yan-lin, LI Yan-ming. Subspecies and in vitro drug sensitivity analysis of Mycobacterium intracellular clinical isolates[J]. Chinese Journal of Antituberculosis, 2021, 43(2): 147-152. doi: 10.3969/j.issn.1000-6621.2021.02.009

Figures/Tables 4

References 31

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引物名称	引物序列(5'~3')	片段长度 (bp)
16S rRNA-F	AGAGTTTGATCMTGGCTCAG	1536
16S rRNA-R	CCGTCAATTCMTTTRAGTTT
rpoB-F	GGCAAGGTCACCCCGAAGGG	764
rpoB-R	AGCGGCTGCTGGGTGATCATC
hsp65-F	ATCGCCAAGGAGATCGAGCT	644
hsp65-R	AAGGTGCCGCGGATCTTGTT
16S-23S rRNA ITS-F	AAGTCGTAACAAGGTARCCG	约380
16S-23S rRNA ITS-R	TCGCCAAGGCATCCACC

药品名称	MIC范围(μg/ml)	耐药临界浓度(μg/ml)
克拉霉素	0.06~64	32
乙胺丁醇	0.5~16	8
利福布汀	0.25~8	4
利福平	0.12~8	8
链霉素	0.5~64	64
阿米卡星	1~64	64
利奈唑胺	1~64	32
莫西沙星	0.12~8	4
多西环素	2~16	-
环丙沙星	0.12~32	-
异烟肼	0.25~8	-
乙硫异烟胺	0.30~20	-
复方磺胺甲噁唑	0.12/238~8/152	-

药品名称	胞内分枝杆菌 (97株)		亚种胞内分枝杆菌 (62株)		副胞内分枝杆菌 (17株)		奇美拉分枝杆菌 (18株)		χ²值	P值
药品名称	耐药株数	耐药率(%)	耐药株数	耐药率(%)	耐药株数	耐药率(%)	耐药株数	耐药率(%)	χ²值	P值
克拉霉素	9	9.28	3	4.84	1	5.88	5	27.78	-	0.023^a
乙胺丁醇	43	44.33	28	45.16	5	29.41	10	55.56	2.470	0.311
利福布汀	15	15.46	8	12.90	5	29.41	2	11.11	-	0.249^a
利福平	41	42.27	23	37.10	9	52.94	9	50.00	1.914	0.402
链霉素	35	36.08	26	41.94	3	17.65	6	33.33	3.485	0.200
阿米卡星	11	11.34	9	14.52	0	0.00	2	11.11	-	0.333^a
利奈唑胺	37	38.14	25	40.32	7	41.18	5	27.78	1.011	0.626
莫西沙星	45	46.39	29	46.77	11	64.71	5	27.78	4.804	0.099