广西壮族自治区耐多药结核分枝杆菌耐药情况与基因型特征分析

doi:10.3969/j.issn.1000-6621.2020.06.008

摘要/Abstract

摘要：

目的了解广西壮族自治区(简称“广西”)耐多药结核分枝杆菌(MDR-MTB)的耐药情况、基因型构成及基因型与耐药的相关性,为耐多药结核病的防控提供理论依据。方法采用连续监测的方法,选取位于广西境内东、西、南、北、中的贵港、百色、崇左、桂林和防城港5个市为监测点,采用随机数字表法抽取5个市中的21个县(市、区),纳入于2016—2017年在当地结核病防治(简称“结防”)机构登记治疗且培养阳性的MTB菌株共1514株,使用WHO推荐的比例法对异烟肼(INH)、利福平(RFP)、乙胺丁醇(EMB)、链霉素(Sm)、氧氟沙星(Ofx)和卡那霉素(Km)进行耐药性检测,最终有51株确定为MDR-MTB菌株。运用熔解曲线间隔区寡核苷酸分型法(McSpoligotyping)对MDR-MTB菌株进行基因分型,将分型结果与SpolDB4.0数据库进行比对。结果 51株MDR-MTB菌株对EMB、Sm、Ofx和Km的耐药率分别为41.18%(21/51)、31.37%(16/51)、9.80%(5/51)和1.96%(1/51)。北京基因型菌株占56.86%(29/51),非北京基因型菌株占43.14%(22/51)。对EMB耐药的菌株中,北京基因型14株,占66.67%(14/21),非北京基因型7株,占33.33%(7/21),差异无统计学意义(χ ²=1.399,P=0.237);对Sm耐药的菌株中,北京基因型11株,占68.75%(11/16),非北京基因型5株,占31.25%(5/16),差异无统计学意义(χ ²=1.343,P=0.246);对Ofx耐药的菌株中,北京基因型9株,占60.00%(9/15),非北京基因型6株,占40.00%(6/15),差异无统计学意义(χ ²=0.085,P=0.770);对Km耐药的菌株中,北京基因型1株,占100.00%(1/1),非北京基因型0株,占0.00%(0/1),差异无统计学意义(P=1.000)。结论应重视广西MDR-MTB菌株对EMB、Sm、Ofx和Km的耐药情况;MDR-MTB菌株主要为北京基因型;北京和非北京基因型对EMB、Sm、Ofx和Km的耐药率未见差异。

关键词: 结核, 抗多种药物性, 基因型, 寡核苷酸序列分析, 数据说明, 统计, 小地区分析

Abstract:

Objective To understand the drug resistance situation, genotype composition and the correlation between genotype and drug-resistant of multidrug-resistant Mycobacterium tuberculosis (MDR-MTB) in Guangxi Zhuang Autonomous Region (referred to as Guangxi), thereby providing a theoretical basis for the prevention and control of MDR-MTB. Methods Using continuous monitoring methods, 5 cities including Guigang, Baise, Chongzuo, Guilin, and Fangchenggang on East, West, South, North, and Central of Guangxi were selected as monitoring points, and random number table method was used to extract 21 counties (cities, districts) from 5 cities. A total of 1514 culture-positive MTB strains which registered and treated in the local tuberculosis prevention and treatment institution were collected from 2016 to 2017. The drug susceptibility test to isoniazid (INH), rifampin (RFP), ethambutol (EMB), streptomycin (Sm), Ofloxacin (Ofx) and kanamycin (Km) was performed using the ratio method recommended by WHO. Finally, 51 strains were identified as MDR-MTB strains. Genotyping was performed by melting curve analysis-based spoligotyping (McSpoligotyping), and then the results were compared with the SpolDB 4.0 database. Results The drug resistance rates to EMB, Sm, Ofx and Km of 51 MDR-MTB strains were 41.18% (21/51), 31.37% (16/51), 9.80% (5/51) and 1.96% (1/51), respectively. Beijing genotype strains accounted for 56.86% (29/51) and non-Beijing genotype strains accounted for 43.14% (22/51). Among the strains resistant to EMB, there were 14 Beijing genotype strains, accounting for 66.67% (14/21) and 7 non-Beijing genotype strains, accounting for 33.33% (7/21), the difference was not statistically significant (χ ²=1.399, P=0.237). Among the strains resistant to Sm, there were 11 Beijing genotype strains, accounting for 68.75% (11/16) and 5 non-Beijing genotype strains, accounting for 31.25% (5/16), the difference was not statistically significant (χ ²=1.343, P=0.246). Among the strains resistant to Ofx, there were 9 Beijing genotype strains, accounting for 60.00% (9/15) and 6 non-Beijing genotype strains, accounting for 40.00%(6/15), the difference was not statistically significant (χ ²=0.085, P=0.770). Among the strains resistant to Km, there was 1 Beijing genotype strain, accounting for 100.00% (1/1), and no non-Beijing genotype strain, accounting for 0.00% (0/1), the difference was not statistically significant (P=1.000). Conclusion We should pay attention to the resistance of MDR-MTB strains to EMB, Sm, Ofx and Km in Guangxi. MDR-MTB strains are mainly Beijing genotype. There are no significant differences in the resistance rates of Beijing and non-Beijing genotypes to EMB, Sm, Ofx and Km.

Key words: Tuberculosis, multidrug-resistant, Gene, Oligonucleotide array sequence analysis, Data interpretation, statistical, Small-area analysis

梁小烟, 林玫, 梁大斌, 蓝如束, 覃慧芳, 叶婧, 黄莉雯. 广西壮族自治区耐多药结核分枝杆菌耐药情况与基因型特征分析[J]. 中国防痨杂志, 2020, 42(6): 578-582. doi: 10.3969/j.issn.1000-6621.2020.06.008

LIANG Xiao-yan, LIN Mei, LIANG Da-bin, LAN Ru-shu, QIN Hui-fang, YE Jing, HUANG Li-wen. Drug resistance and genotypic characteristics of multidrug-resistant Mycobacterium tuberculosis in Guangxi Zhuang Autonomous Region[J]. Chinese Journal of Antituberculosis, 2020, 42(6): 578-582. doi: 10.3969/j.issn.1000-6621.2020.06.008

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参考文献 21

[1]	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.
[2]	董文竹, 王政, 文舒安 , 等. 耐多药及广泛耐药MTB菌株对SQ109的耐药性及其与北京基因型的关系. 中国防痨杂志, 2018,40(5):506-511.
[3]	Pang Y, Zhou Y, Zhao B , et al. Spoligotyping and drug resistance analysis of Mycobacterium tuberculosis strains from national survey in China. PloS One, 2012,7(3):e32976.
[4]	Zeng X, Xu Y, Zhou Y , et al. McSpoligotyping, a One-Step Melting Curve Analysis-Based Protocol for Spoligotyping of Mycobacterium tuberculosis. J Clin Microbiol, 2018,56(8):e00539-18.
[5]	梁大斌, 黄敏莹, 林玫 , 等. 2011—2015年广西肺结核流行情况分析. 应用预防医学, 2017,23(2):133-138.
[6]	罗丹 . 广西耐药结核病流行特征及结核分枝杆菌基因分型的研究. 南宁:广西医科大学, 2013.
[7]	World Health Organization. Global Working Group on Anti-Tuberculosis Drug Resistance Surveillance.Guidelines for surveillance of Drug resistance in tuberculosis. Geneva: World Health Organization, 1997.
[8]	龙茜 . 广西边境与非边境地区结核分枝杆菌耐药及基因型分析. 南宁:广西医科大学, 2019.
[9]	Xu C, Pang Y, Li R , et al. Clinical outcome of multidrug-resistant tuberculosis patients receiving standardized second-line treatment regimen in China. J Infect, 2018,76(4):348-353.
[10]	Wang J, Zhou M, Chen Z , et al. Survival of patients with multidrug-resistant tuberculosis in Central China: a retrospective cohort study. Epidemiol Infect, 2020,148:e50.
[11]	赵冰, 宋媛媛, 逄宇 , 等. 中国耐多药结核分枝杆菌二线抗结核药物敏感性分析. 中国防痨杂志, 2013,35(10):831-834.
[12]	周云, 刘瑞, 杜永国 , 等. 海南省2014—2016年128株耐多药结核分枝杆菌药敏结果. 中国热带医学, 2018,18(6):551-554.
[13]	马丽, 杨建东, 陈阳贵 , 等. 乌鲁木齐市42株耐多药结核病患者耐药情况分析. 中国防痨杂志, 2016,38(10):888-891.
[14]	刘桑, 吕康言, 孟皎 , 等. 360例耐多药肺结核患者耐药情况分析. 中国防痨杂志, 2017,39(8):878-882.
[15]	喻容, 石国民, 马小华 , 等. 2014年长沙市155株耐多药结核分枝杆菌耐药结果分析. 实用预防医学, 2016,23(7):777-779.
[16]	林淑芳, 魏淑贞, 赵永 , 等. 福建省耐多药结核分枝杆菌对吡嗪酰胺的耐药水平与基因型特征分析. 中国防痨杂志, 2018,40(12):1330-1334.
[17]	陈杏 . 中国南方四省结核分枝杆菌分离株基因分型与一线药物耐药性研究. 衡阳:南华大学, 2017.
[18]	石洁, 郑丹薇, 朱岩昆 , 等. 河南省结核菌分离株26位点MIRU-VNTR和Spoligotyping分子分型及耐药分析. 中华疾病控制杂志, 2018,22(11):46-51.
[19]	李妍, 曾小红, 杨健 , 等. 陕西省87株耐药结核分枝杆菌寡核苷酸基因分型和耐药性分析. 现代预防医学, 2018,45(9):149-153.
[20]	Zhao LL, Li MC, Liu HC , et al. Beijing genotype of Mycobacterium tuberculosis is less associated with drug resistance in south China. Int J Antimicrob Agents, 2019,54(6):766-770.
[21]	Liu H, Zhang Y, Liu Z , et al. Associations between Mycobacterium tuberculosis Beijing genotype and drug resistance to four first-line drugs: a survey in China. Front Med, 2018,12(1):92-97. doi: 10.1007/s11684-017-0610-z URL