Identification and drug resistance analysis of non-tuberculous mycobacterial pulmonary disease pathogens in suburban areas of Hangzhou

doi:10.19983/j.issn.2096-8493.20250062

Abstract

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

CLC Number:

R378.91

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

Figures/Tables 2

References 36

[1]	Cowman S, van Ingen J, Griffith DE, et al. Non-tuberculous mycobacterial pulmonary disease. Eur Respir J, 2019, 54(1): 1900250. doi:10.1183/13993003.00250-2019.
[2]	Gu Y, Nie W, Huang H, et al. Non-tuberculous mycobacterial disease: progress and advances in the development of novel candidate and repurposed drugs. Front Cell Infect Microbiol, 2023, 13: 1243457. doi:10.3389/fcimb.2023.1243457.
[3]	刘海灿, 黄明翔, 蒋毅, 等. 福建省肺非结核分枝杆菌临床分离株的菌种鉴定. 中国人兽共患病学报, 2017, 33(5): 389-397. doi:10.3969/j.issn.1002-2694.2017.05.002.
[4]	中华医学会结核病学分会. 非结核分枝杆菌病诊断与治疗指南(2020年版). 中华结核和呼吸杂志, 2020, 43(11): 918-946. doi:10.3760/cma.j.cn112147-20200508-00570.
[5]	牛海军, 李远春, 赵雁林, 等. 河南省南阳市276例非结核分枝杆菌肺病的临床特征及其耐药性分析. 疾病监测, 2021, 36(7): 708-713. doi:10.3784/jbjc.202101290055.
[6]	中国防痨协会. 结核病实验室检验规程. 北京: 人民卫生出版社, 2015.
[7]	Daley CL, Iaccarino JM, Lange C, et al. Treatment of Nontuberculous Mycobacterial Pulmonary Disease: An Official ATS/ERS/ESCMID/IDSA Clinical Practice Guideline. Clin Infect Dis, 2020, 71(4): 905-913. doi:10.1093/cid/ciaa1125. pmid: 32797222
[8]	全国第五次结核病流行病学抽样调查技术指导组,全国第五次结核病流行病学抽样调查办公室. 2010年全国第五次结核病流行病学抽样调查报告. 中国防痨杂志, 2012, 34(8): 485-508.
[9]	Chin KL, Sarmiento ME, Alvarez-Cabrera N, et al. Pulmonary non-tuberculous mycobacterial infections: current state and future management. Eur J Clin Microbiol Infect Dis, 2020, 39(5):799-826. doi:10.1007/s10096-019-03771-0.
[10]	张洁, 苏建荣, 丁北川, 等. 北京地区非结核分枝杆菌菌种分布及耐药性研究. 中华结核和呼吸杂志, 2017, 40(3): 210-214. doi:10.3760/cma.j.issn.1001-0939.2017.03.013.
[11]	洪创跃, 李金莉, 赵广录, 等. 2013—2017年深圳市非结核分枝杆菌流行状况分析. 中国防痨杂志, 2019, 41(5): 529-533. doi:10.3969/j.issn.1000-6621.2019.05.011.
[12]	余旭良, 徐礼锋, 祝进, 等. 99株非结核分枝杆菌菌种鉴定和药敏结果分析. 中国卫生检验杂志, 2015, 25(20):3580-3582. doi:cnki:sun:zwjz.0.2015-20-062.
[13]	廖鑫磊, 王桂荣. 非结核分枝杆菌病分子诊断技术概述. 结核与肺部疾病杂志, 2021, 2(2): 98-101. doi:10.3969/j.issn.2096-8493.2021.02.002.
[14]	潘爱珍, 吴蓓蓓, 柳正卫, 等. 浙江省非结核分枝杆菌病原谱的分析. 中国卫生检验杂志, 2015, 25(24): 4298-4303. doi:cnki:sun:zwjz.0.2015-24-046.
[15]	Zhu Y, Hua W, Liu Z, et al. Identification and characterization of nontuberculous mycobacteria isolated from suspected pulmonary tuberculosis patients in eastern china from 2009 to 2019 using an identification array system. Braz J Infect Dis, 2022, 26(2): 102346. doi:10.1016/j.bjid.2022.102346.
[16]	王少华, 常文静, 苏茹月, 等. 河南省363株非结核分枝杆菌分离鉴定及耐药分析. 中国人兽共患病学报, 2023, 39(2): 124-130. doi:10.3969/j.issn.1002-2694.2022.00.193.
[17]	封彬彬, 金锋, 景辉, 等. 山东地区439株非结核分枝杆菌临床分离株菌种鉴定及耐药情况. 中国防痨杂志, 2021, 43(5): 516-521. doi:10.3969/j.issn.1000-6621.2021.05.019.
[18]	Wang HX, Yue J, Han M, et al. Nontuberculous mycobacteria: susceptibility pattern and prevalence rate in Shanghai from 2005 to 2008. Chin Med J (Engl), 2010, 123(2): 184-187.
[19]	李文彬, 胡培磊, 陈忠南, 等. 湖南省525株非结核分枝杆菌临床分离株的菌种鉴定与流行特征. 中国人兽共患病学报, 2022, 38(5): 417-422. doi:10.3969/j.issn.1002-2694.2022.00.050.
[20]	陈华, 赖晓宇, 陈品儒, 等. 广州地区2018—2021年非结核分枝杆菌临床分离株菌种鉴定结果分析. 实用医学杂志, 2022, 38(17): 2191-2197. doi:10.3969/j.issn.1006-5725.2022.17.014.
[21]	罗春明, 邹桂敏, 刘国标, 等. 广州市2003—2012年非结核分枝杆菌菌种鉴定结果分析. 结核病与肺部健康杂志, 2014, 3(1): 15-20. doi:10.3969/j.issn.2095-3755.2014.01.004.
[22]	Zhang H, Luo M, Zhang K, et al. Species identification and antimicrobial susceptibility testing of non-tuberculous mycobacteria isolated in Chongqing, Southwest China. Epidemiol Infec, 2020, 149: e7. doi:10.1017/S0950268820003088.
[23]	Boyle DP, Zembower TR, Reddy S, et al. Comparison of Clinical Features, Virulence, and Relapse among Mycobacterium avium Complex Species. Am J Respir Crit Care Med, 2015, 191(11): 1310-1317. doi:10.1164/rccm.201501-0067OC.
[24]	Daley CL, Winthrop KL. Mycobacterium avium Complex: Addressing Gaps in Diagnosis and Management. J Infect Dis, 2020, 222(Suppl 4): S199-S211. doi:10.1093/infdis/jiaa354.
[25]	Hoefsloot W, van Ingen J, Andrejak C, et al. The geographic diversity of nontuberculous mycobacteria isolated from pulmonary samples: an NTM-NET collaborative study. Eur Respir J, 2013, 42(6): 1604-1613. doi:10.1183/09031936.00149212. pmid: 23598956
[26]	Lipman M, Cleverley J, Fardon T, et al. Current and future management of non-tuberculous mycobacterial pulmonary disease (NTM-PD) in the UK. BMJ Open Respir Res, 2020, 7(1): e000591. doi:10.1136/bmjresp-2020-000591.
[27]	Porvaznik I, Solovič I, Mokrý J. Non-Tuberculous Mycobacteria: Classification, Diagnostics, and Therapy. Adv Exp Med Biol, 2017, 944:19-25. doi:10.1007/5584_2016_45. pmid: 27826888
[28]	Jeon D. Infection Source and Epidemiology of Nontuberculous Mycobacterial Lung Disease. Tuberc Respir Dis (Seoul), 2019, 82(2): 94-101. doi:10.4046/trd.2018.0026.
[29]	Prasanna AN, Mehra S. Comparative phylogenomics of pathogenic and non-pathogenic Mycobacterium. PLoS One, 2013, 8(8): e71248. doi:10.1371/journal.pone.0071248.
[30]	Prato BD, Altieri AM, Carlucci B, et al. Non-tuberculous mycobacterial pulmonary disease: an Italian National survey. Sarcoidosis Vasc Diffuse Lung Dis, 2018, 35(1): 21-25. doi:10.36141/svdld.v35i1.6979.
[31]	Danley J, Kwait R, Peterson DD, et al. Normal estrogen, but low dehydroepiandrosterone levels, in women with pulmonary Mycobacterium avium complex. A preliminary study. Ann Am Thorac Soc, 2014, 11(6): 908-914. doi:10.1513/AnnalsATS.201312-422OC.
[32]	Uwamino Y, Nishimura T, Sato Y, et al. Low serum estradiol levels are related to Mycobacterium avium complex lung disease: a cross-sectional study. BMC Infect Dis, 2019, 19(1):1055. doi:10.1186/s12879-019-4668-x.
[33]	秦中华, 景晔, 杜岩青, 等. 339株非结核分枝杆菌临床分离株菌种鉴定及耐药性分析. 中国防痨杂志, 2020, 42(6): 630-633. doi:10.3969/j.issn.1000-6621.2020.06.017.
[34]	Huang WC, Yu MC, Huang YW, et al. Identification and drug susceptibility testing for nontuberculous mycobacteria. J Formos Med Assoc, 2020, 119(Suppl 1): S32-S41. doi:10.1016/j.jfma.2020.05.002.
[35]	欧庆芬. 非结核分枝杆菌肺病的CT诊断及鉴别诊断. 结核与肺部疾病杂志, 2024, 5(S): 13-14. doi:10.19983/j.issn.2096-8493.20240357.
[36]	王熠, 陆静, 李苏梅, 等. 84例非结核分枝杆菌肺病患者临床特征及耐药性分析. 结核与肺部疾病杂志, 2021, 2 (3): 223-227. doi:10.3969/j.issn.2096-8493.20210054.

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)