杭州郊县区非结核分枝杆菌肺病病原菌鉴定及其耐药性分析

doi:10.19983/j.issn.2096-8493.20250062

摘要/Abstract

摘要：

目的: 了解近年来杭州郊县农村地区的非结核分枝杆菌(nontuberculous mycobacteria, NTM)肺病的主要流行菌种及其耐药特征。方法: 收集2021年1月至2024年12月杭州8个郊县区结核病定点医院门诊分离的分枝杆菌菌株共3311株,采用对硝基苯甲酸(PNB)和噻吩-2-羧酸肼(TCH)生长试验进行NTM初步鉴定,通过分枝杆菌分子生物学熔解曲线法检测和16S rRNA基因、ITS基因、Hsp65基因的PCR产物测序法对初步获得的NTM进行精确菌种鉴定。采取固体浓度比例法药物敏感性试验(简称“药敏试验”)检测NTM分离菌株对异烟肼、利福平、链霉素、乙胺丁醇、氧氟沙星、左氧氟沙星、莫西沙星和卡那霉素等8种药物的敏感性。结果: 12.4%(409/3311)的分枝杆菌培养阳性标本在PNB/TCH生长试验初筛中获得阳性结果。409例初筛判定为NTM的标本中,22株(5.4%)为结核分枝杆菌,19株(4.6%)为非分枝杆菌属细菌,实际纳入本次研究的NTM肺病患者总数为368例。分子生物学方法鉴定结果显示,368例NTM肺病患者中共鉴定出22种非结核分枝杆菌,其中胞内分枝杆菌占60.6%(223/368),脓肿分枝杆菌占11.7%(43/368),随后依次为堪萨斯分枝杆菌、鸟分枝杆菌、龟分枝杆菌和马赛分枝杆菌,所占比例分别为6.5%(24/368)、5.2%(19/368)、4.6%(17/368)和2.7%(10/368)。368株NTM分离株对异烟肼、利福平、链霉素、乙胺丁醇、氧氟沙星、左氧氟沙星、莫西沙星和卡那霉素的耐药率分别为99.2%(365/368)、82.3%(303/368)、93.5%(344/368)、73.4%(270/368)、87.2%(321/368)、86.1%(317/368)、85.1%(313/368)和80.7%(297/368)。结论: 杭州郊县区NTM肺病致病的优势流行菌种为胞内分枝杆菌,NTM菌株对常规一线及氟喹诺酮类抗结核药物有较高的耐药性。

关键词: 分枝杆菌属, 非典型性细菌, 抗药性, 疾病易感性

Abstract:

Objective: To understand the prevalence and drug resistance of non-tuberculous mycobacteria (NTM) caused lung disease pathogens in suburban counties in Hangzhou in recent years. Methods: PNB/TCH growth tests were used to preliminarily identify 3311 NTM strains isolated from mycobacterium-positive culture samples from patients visiting tuberculosis designated hospitals in 8 suburban counties and districts in Hangzhou from January 2021 to December 2024. The species of NTM isolates were confirmed using mycobacterium molecular linear probe method and sequencing of PCR product of 16S rRNA, ITS and Hsp65 gene. Concentration proportional method was applied to detect the susceptibility of NTM isolates to isoniazid, rifampicin, streptomycin, ethambutol, ofloxacin, levofloxacin, moxifloxacin, and kanamycin. Results: PNB/TCH growth tests showed that 12.4% (409/3311) of the 3311 mycobacterium-positive culture samples were positive. Out of these 409 specimens preliminarily identified as NTM, 22 (5.4%) strains were Mycobacterium tuberculosis, 19 (4.6%) strains were non mycobacterial. The total number of NTM patients included in this study was 368. The mycobacterium molecular linear probe hybridization and PCR product sequencing confirmed that among these 368 NTM samples, 22 types of nontuberculous mycobacteria were identified, 60.6%(223/368) were identified as Mycobacterium intracellulare, followed by Mycobacterium abscessus (11.7%, 43/368), Mycobacterium kansasii (6.5%, 24/368), Mycobacterium avium (5.2%, 19/368), Mycobacterium chelonei (4.6%, 17/368) and Mycobacterium marseillense (2.7%, 10/368). The resistance rates of 368 NTM isolates to isoniazid, rifampicin, streptomycin, ethambutol, ofloxacin, levofloxacin, moxifloxacin, and kanamycin were 99.2% (365/368), 82.3% (303/368), 93.5% (344/368), 73.4% (270/368), 87.2% (321/368), 86.1% (317/368), 85.1% (313/368), and 80.7% (297/368), respectively. Conclusion: Mycobacterium intracellulare is the predominant NTM species in Hangzhou suburban areas. Moreover, NTM isolates have high resistance against commonly used antituberculosis drugs.

Key words: Mycobacterium, Atypical bacterial forms, Drug resistance, Disease susceptibility

中图分类号:

R378.91

吴亦斐, 李清春, 贾庆军, 黄银燕, 白雪欣, 程庆林. 杭州郊县区非结核分枝杆菌肺病病原菌鉴定及其耐药性分析[J]. 结核与肺部疾病杂志, 2025, 6(4): 420-425. doi: 10.19983/j.issn.2096-8493.20250062

Wu Yifei, Li Qingchun, Jia Qingjun, Huang Yinyan, Bai Xuexin, Cheng Qinglin. Identification and drug resistance analysis of non-tuberculous mycobacterial pulmonary disease pathogens in suburban areas of Hangzhou[J]. Journal of Tuberculosis and Lung Disease, 2025, 6(4): 420-425. doi: 10.19983/j.issn.2096-8493.20250062

图/表 2

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表2

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年龄组(岁)	男性	女性	合计
≤19	2(0.5)	3(0.8)	5(1.4)
20~29	5(1.4)	4(1.1)	9(2.5)
30~39	11(3.0)	2(0.5)	13(3.5)
40~49	13(3.5)	7(1.9)	20(5.4)
50~59	21(5.7)	31(8.4)	52(14.1)
60~69	61(16.6)	34(9.2)	95(25.8)
70~79	82(22.3)	35(9.5)	117(31.8)
≥80	35(9.5)	22(6.0)	57(15.5)
合计	230(62.5)	138(37.5)	368(100.0)

非结核分枝杆菌	株数	异烟肼	利福平	链霉素	乙胺丁醇	氧氟沙星	左氧氟沙星	莫西沙星	卡那霉素
胞内分枝杆菌	223	220(98.7)	190(85.2)	210(94.2)	153(68.6)	200(89.7)	201(90.1)	198(88.8)	189(84.8)
脓肿分枝杆菌	43	43(100.0)	38(88.4)	39(90.7)	37(86.0)	35(81.4)	33(76.7)	33(76.7)	34(79.1)
堪萨斯分枝杆菌	24	24(100.0)	13(54.2)	22(91.7)	11(45.8)	15(62.5)	13(54.2)	13(54.2)	14(58.3)
鸟分枝杆菌	19	19(100.0)	14(73.7)	17(89.5)	17(89.5)	18(94.7)	18(94.7)	18(94.7)	13(68.4)
龟分枝杆菌	17	17(100.0)	11(64.7)	16(94.1)	15(88.2)	15(88.2)	15(88.2)	15(88.2)	15(88.2)
马赛分枝杆菌	10	10(100.0)	9(90.0)	10(100.0)	7(70.0)	10(100.0)	9(90.0)	8(80.0)	8(80.0)
哥伦比亚分枝杆菌	5	5(5/5)	4(4/5)	4(4/5)	5(5/5)	5(5/5)	5(5/5)	5(5/5)	5(5/5)
奇美拉分枝杆菌	4	4(4/4)	4(4/4)	4(4/4)	4(4/4)	4(4/4)	4(4/4)	4(4/4)	3(75.0)
蒂莫内分枝杆菌	3	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	2(66.7)
蟾蜍分枝杆菌	3	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)
偶然分枝杆菌	3	3(3/3)	1(33.3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)	3(3/3)
首尔分枝杆菌	2	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	0(0.0)
M.persicum	2	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)
M.sherrisii	2	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)	2(2/2)
其他^a	8	8(/8)	7(7/8)	7(7/8)	6(6/8)	4(4/8)	4(4/8)	4(4/8)	4(4/8)
合计	368	365(99.2)	303(82.3)	344(93.5)	270(73.4)	321(87.2)	317(86.1)	313(85.1)	297(80.7)