17种抗生素对北京地区快速生长分枝杆菌体外抑菌效果分析

doi:10.19982/j.issn.1000-6621.20240167

摘要/Abstract

摘要：

目的: 分析17种抗生素对北京地区快速生长分枝杆菌(rapidly growing mycobacteria, RGM)临床分离株的抑菌效果,为RGM引起的非结核分枝杆菌病的临床治疗提供可靠的理论依据。方法: 收集北京市疾病预防控制中心结核病实验室2016年1月至2020年12月五年间分离保存的130株非结核分枝杆菌菌株,对其进行菌种鉴定。对鉴定出的RGM临床分离株和相应标准株复苏后,采用微量肉汤稀释法进行17种药物(阿米卡星、环丙沙星、磺胺甲噁唑-甲氧苄啶、克拉霉素、利奈唑胺、莫西沙星、多西环素、米诺环素、头孢西丁、替加环素、妥布霉素、亚胺培南、美罗培南、阿莫西林/克拉维酸、贝达喹啉、德拉马尼和氯法齐明)的敏感性试验,记录其最低抑菌浓度(MIC)及抑制50%或90%受试菌的MIC值(MIC₉₀和MIC₅₀)。结果: 经菌种鉴定且复苏成功的RGM为37株,包括19株脓肿分枝杆菌、17株偶发分枝杆菌和1株龟分枝杆菌。已明确耐药折点的14种抗生素中,所检测的3种RGM临床分离株对替加环素均敏感,龟分枝杆菌、脓肿分枝杆菌临床分离株对克拉霉素均敏感,脓肿分枝杆菌临床分离株对利奈唑胺敏感。另外,偶发分枝杆菌临床分离株对阿米卡星、环丙沙星、磺胺甲噁唑-甲氧苄啶、莫西沙星对均敏感。脓肿分枝杆菌临床分离株对阿米卡星的敏感率为73.7%(14/19),偶发分枝杆菌临床分离株对利奈唑胺、美罗培南敏感率分别为82.4%(14/17)、88.2%(15/17)。所检测的19株脓肿分枝杆菌对多西环素、阿莫西林/克拉维酸均耐药,偶发分枝杆菌临床分离株对妥布霉素均耐药。此外,贝达喹啉和氯法齐明对RGM临床分离株的MIC₉₀均小于药敏板最低检测限值,而德拉马尼的MIC₉₀均高于药敏板最大检测限值。结论: 替加环素、阿米卡星、利奈唑胺、贝达喹啉和氯法齐明对RGM均具有较好的体外抑菌活性,而多西环素、阿莫西林/克拉维酸、德拉马尼几乎没有体外抑菌活性。非结核分枝杆菌种间耐药谱差异较大,建议用药前进行菌种鉴定和药物敏感性试验,以选择合适的抗菌药物。

关键词: 分枝杆菌,非典型性, 快速生长分枝杆菌, 微生物敏感性试验, 评价研究

Abstract:

Objective: To analyze the antibacterial effects of 17 antibiotics on clinical isolates of rapidly growing mycobacteria (RGM) in Beijing, and to provide a reliable theoretical basis for the clinical treatment of nontuberculosis mycobacteriosis caused by RGM. Methods: A total of 130 nontuberculous mycobacteria (NTM) strains isolated from the Tuberculosis Laboratory of the Beijing Center for Disease Control and Prevention between January 2016 and December 2020 were collected for strain identification. After reviving the RGM clinical isolates and corresponding standard strains, sensitivity tests were conducted on 17 antibiotics (amikacin, ciprofloxacin, compound sulfamethoxazole, clarithromycin, linezolid, moxifloxacin, doxycycline, minocycline, cefoxitin, tigecycline, tobramycin, imipenem, meropenem, amoxicillin/clavulanic acid, bedaquiline, delamanid, and clofazimine) using the micro broth dilution method. The minimum inhibitory concentration (MIC) and the MIC values required to inhibit 50% (MIC₅₀) or 90% (MIC₉₀) of the tested bacteria were recorded. Results: After strain identification and successful recovery, 37 strains of RGM were identified, including 19 Mycobacterium (M.) abscesses, 17 M.fortuitum, and 1 M.chelonae. Among the 14 antibiotics with established resistance breakpoints, all RGM clinical isolates were found to be sensitive to tigecycline. Clinical isolates of M.chelonae and M.abscessus were sensitive to clarithromycin, and M.abscesses were also sensitive to linezolid. All M.fortuitum clinical isolates were sensitive to amikacin, ciprofloxacin, compound sulfamethoxazole, and moxifloxacin. Additionally, the sensitivity rate of M.abscessus to amikacin was 73.7% (14/19), while the sensitivity rates of M.fortuitum to linezolid and meropenem were 82.4% (14/17) and 88.2% (15/17), respectively. All M.abscessus isolates were resistant to doxycycline, amoxicillin/clavulanic acid, while all M.fortuitum isolates were resistant to tobramycin. Additionally, the MIC₉₀ of bedaquiline and clofazimine for RGM clinical isolates were both lower than the minimum detection limit of the drug sensitivity plate, whereas the MIC₉₀ of delamanid was higher than the maximum detection limit of the drug sensitivity plate. Conclusion: Tigecycline, amikacin, linezolid, betaquiline, and clofazimine all exhibit good in vitro antibacterial activity against RGM, while doxycycline, amoxicillin/clavulanic acid, and delamanid show almost no in vitro antibacterial activity. There is a significant difference in the drug resistance spectrum between NTM species. Therefore, it is recommended to conduct species identification and drug sensitivity testing before treatment to select appropriate antibiotics.

Key words: Mycobacteria,atypical, Rapidly growing mycobacteria, Microbial sensitivity tests, Evaluation studies

中图分类号:

陈双双, 田丽丽, 王嫩寒, 杨新宇, 赵琰枫, 李传友, 代小伟. 17种抗生素对北京地区快速生长分枝杆菌体外抑菌效果分析[J]. 中国防痨杂志, 2024, 46(9): 1056-1062. doi: 10.19982/j.issn.1000-6621.20240167

Chen Shuangshuang, Tian Lili, Wang Nenhan, Yang Xinyu, Zhao Yanfeng, Li Chuanyou, Dai Xiaowei. Analysis of in vitro antibacterial effects of 17 antibiotics against rapidly growing mycobacteria in the Beijing area[J]. Chinese Journal of Antituberculosis, 2024, 46(9): 1056-1062. doi: 10.19982/j.issn.1000-6621.20240167

图/表 4

表1

表2

表3

脓肿分枝杆菌菌株的药物敏感性试验结果

抗生素	标准株浓度 (μg/ml)	MIC^b(μg/ml)			菌株数(构成比,%)
抗生素	标准株浓度 (μg/ml)	MIC范围	$MIC 50 c$	$MIC 90 c$	敏感	中介	耐药
阿米卡星	16	4~32	16	32	14(73.7)	5(26.3)	0(0.0)
环丙沙星	1	0.5~>4	2	>4	6(31.6)	9(47.4)	4(21.0)
磺胺甲噁唑- 甲氧苄啶	1/19	0.25/4.8~>8/152	4/76	8/152	5(26.3)	-	14(73.7)
克拉霉素	<1	<1	<1	<1	19(100.0)	0(0.0)	0(0.0)
利奈唑胺	2	1~8	2	8	19(100.0)	0(0.0)	0(0.0)
莫西沙星	<0.25	0.25~>8	2	8	4(21.1)	7(36.8)	8(42.1)
多西环素	>16	16~>16	>16	>16	0(0.0)	0(0.0)	19(100.0)
米诺环素	>8	1~>8	8	>8	3(15.8)	5(26.3)	11(57.9)
头孢西丁	>128	16~64	16	32	11(57.9)	8(42.1)	0(0.0)
替加环素^a	0.5	<0.25~2	<0.25	0.5	19(100.0)	-	0(0.0)
妥布霉素	<1	<1~16	2	4	13(68.4)	5(26.3)	1(5.3)
亚胺培南	64	2~32	8	32	3(15.8)	14(73.7)	2(10.5)
美罗培南	32	2~32	16	32	1(5.3)	10(52.6)	8(42.1)
阿莫西林/克拉维酸	>64/32	>64/32	>64/32	>64/32	0(0.0)	0(0.0)	19(100.0)
贝达喹啉	<0.125	<0.125	<0.125	<0.125	-	-	-
德拉马尼	>0.5	0.5~>0.5	0.5	>0.5	-	-	-
氯法齐明	<0.125	<0.125	<0.125	<0.125	-	-	-

表3

表4

偶发分枝杆菌菌株的药物敏感性试验结果

抗生素	标准株浓度 (μg/ml)	MIC^b(μg/ml)			菌株数(构成比,%)
抗生素	标准株浓度 (μg/ml)	MIC范围	$MIC 50 c$	$MIC 90 c$	敏感	中介	耐药
阿米卡星	<2	<2~4	<2	<2	17(100.0)	0(0.0)	0(0.0)
环丙沙星	<0.12	<0.12~1	<0.12	1	17(100.0)	0(0.0)	0(0.0)
磺胺甲噁唑- 甲氧苄啶	1/19	<0.25/4.8~2/38	1/19	2/38	17(100.0)	-	0(0.0)
克拉霉素	8	<1~32	4	32	8(47.1)	3(17.6)	6(35.3)
利奈唑胺	8	1~>32	8	8	14(82.3)	1(5.9)	2(11.8)
莫西沙星	<0.25	<0.25~1	<0.25	<0.25	17(100.0)	0(0.0)	0(0.0)
多西环素	<0.25	<0.25~>16	>16	>16	5(29.4)	0(0.0)	12(70.6)
米诺环素	<0.5	<0.5~>8	8	>8	5(29.4)	0(0.0)	12(70.6)
头孢西丁	64	16~128	32	64	3(17.6)	13(76.5)	1(5.9)
替加环素^a	<0.25	<0.25~0.5	<0.25	0.5	17(100.0)	-	0(0.0)
妥布霉素	16	8~>16	16	16	0(0.0)	0(0.0)	17(100.0)
亚胺培南	8	<2~8	4	8	9(52.9)	8(47.1)	0(0.0)
美罗培南	4	<2~8	4	8	15(88.2)	2(11.8)	0(0.0)
阿莫西林/克拉维酸	8/4	8/4~>64/32	>64/32	>64/32	1(5.9)	0(0.0)	16(94.1)
贝达喹啉	<0.125	<0.125~0.25	<0.125	<0.125	-	-	-
德拉马尼	>0.5	>0.5	>0.5	>0.5	-	-	-
氯法齐明	<0.125	<0.125	<0.125	<0.125	-	-	-

表4

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抗生素	浓度范围	MIC临界浓度(μg/ml)^a
抗生素	浓度范围	敏感	中介	耐药
阿米卡星	2~128	≤16	32	≥64
环丙沙星	0.12~4	≤1	2	≥4
磺胺甲噁唑-甲氧苄啶	0.25/4.8~8/152	≤2/38	-	≥4/76
克拉霉素	1~64	≤2	4	≥8
利奈唑胺	0.125~32	≤8	16	≥32
莫西沙星	0.25~8	≤1	2	≥4
多西环素	0.25~16	≤1	2~4	≥8
米诺环素	0.5~8	≤1	2~4	≥8
头孢西丁	4~128	≤16	32~64	≥128
替加环素^b	0.25~8	≤4	-	>4
妥布霉素	1~16	≤2	4	≥8
亚胺培南	2~64	≤4	8~16	≥32
美罗培南	2~64	≤4	8~16	≥32
阿莫西林/克拉维酸	4/2~64/32	≤8/4	16/8	≥32/16
贝达喹啉	0.125~8	-	-	-
德拉马尼	0.006~0.5	-	-	-
氯法齐明	0.125~8	-	-	-