中国异烟肼耐药结核分枝杆菌基因突变特征分析

doi:10.19983/j.issn.2096-8493.2024118

摘要/Abstract

摘要：

目的：总结异烟肼(INH)耐药结核分枝杆菌菌株耐药基因类型、突变位点、地域差异等方面特征,为结核病的分子生物学诊断、临床用药、耐药基因的新发现等研究提供理论依据。方法：该研究在中国知网、维普、万方数据库中以“异烟肼耐药并且突变并且结核”为检索式检索有关中国结核分枝杆菌耐INH基因突变的研究文献,在PubMed以“Isoniazid OR INH” AND “mutation” AND “Mycobacterium tuberculosis” AND “China”为检索式检索中国相关英文文献,初步检索145篇文献,实际纳入文献22篇,纳入分析菌株12923株,获得INH耐药菌株2824株。以2019年为时间节点,将国内关于INH耐药基因突变信息分为两部分进行研究,对耐药基因类型、突变位点、耐药菌株地域差异等信息进行分析。结果：2019年后,96.18%(2716/2824)检测到INH耐药基因突变,常见突变基因为katG(315位点),占74.49%(1965/2638),以AGC→ACC突变为主,占52.26%(1027/1965);inhA基因-15位点突变最常发生,占13.72%(362/2638),以C→T为主,占89.23%(323/362);1.97%(52/2638)fabG1发生突变。联合突变以katG+inhA突变为主,占2.20%(58/2638)。结论：中国耐INH结核分枝杆菌的耐药基因中,katG、inhA、fabG1基因突变常发,突变位点以katG315、inhA15为主,联合突变以katG+inhA突变为主。

关键词: 分枝杆菌,结核, 抗药性, 突变, 异烟肼

Abstract:

Objective: To summarize the mutation characteristics of isoniazid (INH)-resistant strains in terms of drug-resistant gene types, mutation sites and geographical differences, to provide a theoretical basis for the molecular diagnosis of anti-tuberculosis, clinical use of medicines, and the discovery of new drug-resistant genes. Methods: We searched for mutations in INH-resistant genes of Mycobacterium tuberculosis (M.tuberculosis) in China in China Knowledge Network (CNN), Weipu, and Wanfang database with the search terms of “isoniazid resistance” AND “mutation” AND “tuberculosis”, and searched for literature in English with the search terms of “Isoniazid OR INH” AND “mutation” AND “Mycobacterium tuberculosis” AND “China” in PubMed. 145 articles were initially identified, 22 articles were included, 12923 strains were included in the analysis, and 2824 INH-resistant strains were obtained. Taking year 2019 as a time point, the domestic information on INH drug-resistant gene mutations was divided into two parts for studying, then information on drug-resistant gene types, mutation sites, and geographic differences of drug-resistant strains was analyzed in detail. Results: Mutations in INH resistance genes were detected in 96.18% (2716/2824) of strains. The common mutated genes were katG (locus 315), accounting for 74.49% (1965/2638), for which AGC→ACC mutations were predominantly found, accounting for 52.26% (1027/1965). Mutations at locus -15 of the inhA gene were most frequent, accounting for 13.72% (362/2638) with C→T predominating at 89.23% (323/362). fabG1 mutation accounted for 1.97% (52/2638), and the most frequent combined mutation was katG+inhA, accounting for 2.20% (58/2638). Conclusion: Among drug-resistant genes of INH-resistant M.tuberculosis in China, mutations in katG, inhA, and fabG1 gene occured frequently. Mutation loci were dominated by katG315 and inhA15, the most frequent combined mutation was katG+inhA.

Key words: Mycobacterium tuberculosis, Drug resistance, Mutation, Isoniazid

中图分类号:

R183.3

徐雁南, 方梓昊, 赵文丽, 郑佳雄, 刘苏洋, 林健雄, 纪丽微, 常巧呈. 中国异烟肼耐药结核分枝杆菌基因突变特征分析[J]. 结核与肺部疾病杂志, 2025, 6(1): 14-21. doi: 10.19983/j.issn.2096-8493.2024118

Xu Yannan, Fang Zihao, Zhao Wenli, Zheng Jiaxiong, Liu Suyang, Lin Jianxiong, Ji Liwei, Chang Qiaocheng. Characterisation of isoniazid-resistant Mycobacterium tuberculosis mutations in China[J]. Journal of Tuberculosis and Lung Disease, 2025, 6(1): 14-21. doi: 10.19983/j.issn.2096-8493.2024118

图/表 7

图1

表1

表2

表3

表4

表5

结核分枝杆菌异烟肼耐药基因突变情况

突变基因	位点/密码子	氨基酸改变	菌株数(株)	突变基因	位点/密码子	氨基酸改变	菌株数(株)
katG	129	Phe→Ser	1	inhA	-8T→A	-	2
katG	142GAC→GGC	Asn→Asp	1	inhA	-8T→C	-	2
katG	190CAG→UAG	Gln→SC	1	inhA	-9G→A	-	1
katG	232CCG→ACG	Pro→Thr	1	inhA	-15C→T	-	323
katG	285GGC→TGC	GIy→Cys	1	inhA	24C→T	-	1
katG	289GAG→GGG	GIu→Gly	1	inhA	93G→T	-	1
katG	300TGG→AGG	Trp→Arg	1	inhA	154G→A	-	1
katG	315AGC→ACC	Ser→Thr	1027	inhA	94UCG→GCG	Ser→Ala	2
katG	315AGC→AAC	Ser→Asn	51	inhA	142GGU→AGU	Gly→Ser	1
katG	315AGC→ACA	Ser→Thr	1	inhA	194AUG→ACC	Met→Thr	1
katG	315AGC→AGA	Ser→Arg	2	inhA	194AUC→ACC	Ile→Thr	1
katG	315AGC→AUC	Ser→Lie	3	inhA	其他^b	-	28
katG	315AGC→CGC	Ser→Arg	1	ahpC	-47插入T	-	3
katG	315AGC→GGC	Ser→Gly	2	ahpC	-47G→A	-	1
katG	315AGC→AGU	Ser→Ser	2	ahpC	-48G→A	-	6
katG	315	Ser→Thr	486	ahpC	-52C→T	-	11
katG	315	Ser→Asn	22	ahpC	-52C→A	-	2
katG	315缺失	-	4	ahpC	-54C→T	-	4
katG	316GGC→CGC	Gly→Arg	1	ahpC	-57C→T	-	1
katG	317	Ile→Val	1	ahpC	68C→T	-	2
katG	328UGG→UCG	Trp→Ser	1	ahpC	-74G→A	-	2
katG	333CUC→CCC	Leu→Arg	1	ahpC	-81C→T	-	5
katG	335AUC→ACC	Ile→Thr	2	ahpC	481G→A	-	6
katG	419GAC→CAC	Asp→His	180	ahpC	31CCC→CCU	Pro→Pro	1
katG	448GAC→GGC	Asp→Gly	1	ahpC	106UUA→GUA	Leu→Val	1
katG	461CAG→CCG	GIn→Pro	2	ahpC	其他^c	-	15
katG	463CGG→CUG	Arg→Leu	189	ndh	285UUG→CUG	Leu→Leu	2
katG	735GAC→GCC	Asp→AIa	1	ndh	329UGG→UAG	Trp→stop	1
katG	1317AUC→GUC	Ile→Val	1	ndh	339GGG→GCG	GIy→AIa	1
katG	AAC→GAC	Asn→Phe	1	ndh	364UAC→UAU	Tyr→Tyr	1
katG	129	Asp→Ser	1	ndh	399CGA→CGC	Arg→Arg	1
katG	138	GIn→His	1	kasA	121AGG-AAG	Arg→Lys	1
katG	09	AlaI→Val	1	kasA	164GUG→GUA	Val→Val	2
katG	缺失	-	1	kasA	252GCA→GUA	AIa→Val	1
katG	插入突变CG	-	1	kasA	312GGC→AGC	GIy→Ser	3
katG	其他^a	-	170	kasA	404位后插入C	-	2
katG+ahpC	-	-	16	kasA	其他^d	-	3
katG+inhA	-	-	58	katG+katG	-	-	10
其他组合^e	-	-	76	其他^f	-	-	52

表5

表6

参考文献 38

[1]	贾平平, 余利岩, 岑山. 结核分枝杆菌耐一线耐药基因突变及分子机制研究. 中国抗生素杂志, 2011, 36(7): 487-490. doi:10.3969/j.issn.1006-740X.2012.01.005.
[2]	张晨曦, 刘鼎阔. 细菌群集运动特性的研究进展. 生物工程学报, 2023, 39(8):3188-3203. doi:10.13345/j.cjb.220892.
[3]	Holzheimer M, Buter J, Minnaard AJ. Chemical Synthesis of Cell Wall Constituents of Mycobacterium tuberculosis. Chem Rev, 2021, 121(15): 9554-9643. doi:10.1021/acs.chemrev. pmid: 34190544
[4]	葛赛, 宋欣怡, 姜惠跃, 等. 结核分枝杆菌药物作用机理及耐药机制研究进展. 海南医学院学报, 2023, 29(24): 1904-1912. doi:10.13210/j.cnki.jhmu.20221128.002.
[5]	董思佳, 王毳. 谈耐多药结核病的防控进展与策略. 中国热带医学, 2021, 21(3):282-286. doi:10.13604/j.cnki.46-1064/r.2021.03.19.
[6]	龚兰, 牛群, 王楠, 等. 结核分枝杆菌异烟肼耐药基因突变与其他抗结核药物耐药的相关性. 实用医学杂志, 2023, 39(11): 1353-1358. doi:10.3969/j.issn.1006-5725.2023.11.006.
[7]	World Health Organization. WHO global tuberculosis report. Geneva: World Health Organization, 2019.
[8]	Gygli SM, Borrell S, Trauner A, et al. Antimicrobial resis-tance in Mycobacterium tuberculosis: mechanistic and evolutionary perspectives. FEMS Microbiol Rev, 2017, 41(3):354-373. doi:10.1093/femsre/fux011.
[9]	Dulberger CL, Rubin EJ, Boutte CC. The mycobacterial cell envelope-a moving target. Nat Rev Microbiol, 2020, 18(1):47-59. doi:10.1038/s41579-019-0273-7. pmid: 31728063
[10]	田丽, 周伟, 黄星, 等. 中国异烟肼耐药结核分枝杆菌基因突变特征分析. 中国防痨杂志, 2022, 44(4):354-361. doi:10.19982/j.issn.1000-6621.20210573.
[11]	杨映晖, 伍定辉, 苏伟明, 等. 基因芯片法在结核分枝杆菌耐药基因快速诊断中的应用价值. 中国防痨杂志, 2020, 42(11):1191-1195. doi:10.3969/j.issn.1000-6621.2020.11.009.
[12]	王嫩寒, 赵琰枫, 田丽丽, 等. 荧光PCR探针熔解曲线技术检测痰标本中结核分枝杆菌异烟肼耐药性的研究. 实用医学杂志, 2023, 39(20):2664-2670. doi:10.19982/j.issn.1000-6621.20230086.
[13]	吴联朋, 徐克, 何贵清. 温州市耐多药/利福平耐药结核分枝杆菌耐药基因突变及特征分析. 温州医科大学学报, 2023, 53(4):305-310. doi:10.3784/j.issn.1003-9961.2014.04.014.
[14]	蔡莺莺, 石庆新, 周秋菊, 等. 台州地区结核分枝杆菌利福平和异烟肼耐药基因的流行病学调查. 中国现代医生, 2021, 59(20):146-149.
[15]	赵竟男, 江友桥, 伍文华, 等. 江西省3个监测点结核分枝杆菌耐药性及耐药基因检测分析. 疾病监测, 2022, 37(1):127-131. doi:10.3784/jbjc.202107140397.
[16]	徐峰, 饶跃峰, 张幸国, 等. 宁波地区结核分枝杆菌基因突变位点确证及与异烟肼、利福平耐药关系研究. 中国现代应用药学, 2020, 37(20):2511-2515. doi:10.13748/j.cnki.issn1007-7693.2020.20.014.
[17]	王伟, 李静, 郭琪, 等. 耐异烟肼结核分枝杆菌的基因突变类型及表型异质性研究. 中国卫生检验杂志, 2021, 31(22):2744-2746.
[18]	朱大冕, 刘文果, 沈静, 等. 重庆地区结核分枝杆菌异烟肼和丙硫异烟胺耐药及其交叉耐药相关基因突变研究. 中国人兽共患病学报, 2022, 38(5):405-409. doi:10.3969/j.issn.1002-2694.2022.00.047.
[19]	王菊红, 穆丽平. 耐多药结核分枝杆菌异烟肼和利福平耐药相关基因突变特征观察. 内蒙古医学杂志, 2022, 54(8):988-990. doi:10.16096/J.cnki.nmgyxzz.2022.54.08.026.
[20]	杨燕, 张向荣. 结核分枝杆菌异烟肼耐药基因突变与耐药水平的相关性研究. 南京医科大学学报(自然科学版), 2023, 43(3):392-396.
[21]	Wang L, Yang J, Chen L, et al. Whole-genome sequencing of Mycobacterium tuberculosis for prediction of drug resistance. Epidemiol Infect, 2022, 150:22. doi:10.1016/s1473-3099(15)00062-6.
[22]	秦翊翔, 蓝如束, 覃慧芳, 等. 广西壮族自治区结核分枝杆菌异烟肼与利福平耐药基因联合突变特征分析. 疾病监测, 2022, 37(6):787-791. doi:10.3784/jbjc.202112120636.
[23]	陈连勇, 茹浩浩, 杨星, 等. 云南省异烟肼耐药结核分枝杆菌katG和inhA基因突变特征. 昆明医科大学学报, 2022, 43(8):28-33. doi:10.12259/j.issn.2095-610X.S20220803.
[24]	赵刚, 贾庆军, 吴亦斐, 等. 浙江地区耐多药结核分枝杆菌基因突变特征分析. 中华疾病控制杂志, 2021, 25(1):66-71. doi:10.16462/j.cnki.zhjbkz.2021.01.013.
[25]	文书, 林璋礼, 刘丁发, 等. 耐异烟肼结核分枝杆菌及其katG、inhA基因的突变. 中国热带医学, 2019, 19(8):723-726. doi:10.13604/j.cnki.46-1064/r.2019.08.03.
[26]	许河南, 杨红慧, 赖聪娟, 等. 浙江省丽水市结核分枝杆菌临床分离株耐药性及基因突变情况. 疾病监测, 2023, 38(1):51-56. doi:10.3784/jbjc.202205260243.
[27]	蒋燕成, 张建明, 陈紫萱, 等. 结核分枝杆菌耐药基因突变特征及与耐药水平关系的研究. 检验医学与临床, 2023, 20(15):2145-2148. doi:10.3969/j.issn.1672-9455.2023.15.001.
[28]	穆成, 孙蕊, 王志锐, 等. 基于全基因组测序的72株耐多药结核分枝杆菌耐药基因突变特征分析. 中国热带医学, 2023, 23(7):725-729,741. doi:10.13604/j.cnki.46-1064/r.2023.07.09.
[29]	邓丽, 王楠, 雷杰, 等. 华南地区380株结核分枝杆菌异烟肼耐药基因突变情况分析. 国际医药卫生导报, 2023, 29(5):614-618. doi:10.3760/cma.j.issn.1007-1245.2023.05.005.
[30]	Islam MM, Tan Y, Hameed HMA, et al. Detection of novel mutations associated with independent resistance and cross-resistance to isoniazid and prothionamide in Mycobacterium tuberculosis clinical isolates. Clin Microbiol Infect, 2019, 25(8):1041.e1-1041.e7. doi:10.1016/j.cmi.2018.12.008.
[31]	Zhdanova S, Jiao WW, Sinkov V, et al. Insight into Population Structure and Drug Resistance of Pediatric Tuberculosis Strains from China and Russia Gained through Whole-Genome Sequencing. Int J Mol Sci, 2023, 24(12):10302. doi:10.3390/ijms241210302.
[32]	Jia H, Xu Y, Sun Z. Analysis on Drug-Resistance-Associated Mutations among Multidrug-Resistant Mycobacterium tuberculosis Isolates in China. Antibiotics (Basel), 2021, 10(11):1367. doi:10.3390/antibiotics10111367.
[33]	罗红丽, 庞蕾, 谢建平. 分枝杆菌枝菌酸合成及其调控. 微生物学报, 2012, 52(2):146-151. doi:10.13343/j.cnki.wsxb.2012.02.011.
[34]	Oudghiri A, Karimi H, Chetioui F, et al. Molecular characterization of mutations associated with resistance to second-line tuberculosis drug among multidrug-resistant tuberculosis patients from high prevalence tuberculosis city in Morocco. BMC Infect Dis, 2018, 18(1):98. doi:10.1186/s12879-018-3009-9. pmid: 29486710
[35]	Chiaradia L, Lefebvre C, Parra J, et al. Dissecting the mycobacterial cell envelope and defining the composition of the native mycomembrane. Sci Rep, 2017, 7(1):12807. doi:10.1038/s41598-017-12718-4. pmid: 28993692
[36]	Marrakchi H, Lanéelle MA, Daffé M. Mycolic acids: structures, biosynthesis, and beyond. Chem Biol, 2014, 21(1):67-85. doi:10.1016/j.chembiol.2013.11.011.
[37]	Cole S, Brosch R, Parkhill J, et al. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature, 1998, 393(6685):537-544. doi:10.1038/31159.
[38]	石大伟. 河南省耐多药结核分枝杆菌耐药相关基因的分子流行病学研究. 郑州:中国医学科学院病原生物学研究所, 2011.

序号	文献来源	收集菌株年份	结核分枝杆菌数(株)	耐INH菌株数(株)	分子生物学检测技术
1	杨映晖等^[11]	2017—2018	411	138^a	基因芯片法、GeneXpert^c
2	王嫩寒等^[12]	2021—2022	157	35^b	MeltPro技术、Hain试验^d
3	吴联朋等^[13]	2021—2022	54	45	WGS
4	蔡莺莺等^[14]	2015—2020	482	127	芯片杂交法
5	赵竟男等^[15]	2019	250	34	PCR-线性杂交酶法
6	徐峰等^[16]	2017—2018	2455	84	PCR扩增
7	王伟等^[17]	2016—2020	199	94	Xpert MTB/RIF Ultra
8	朱大冕等^[18]	2014—2019	238	233	PCR扩增
9	王菊红和穆丽平^[19]	2019—2020	78	70	实时荧光定量PCR
10	杨燕和张向荣^[20]	2019—2021	1712	404	荧光PCR
11	Wang等^[21]	2017—2018	137	81	WGS
12	秦翊翔等^[22]	2017—2018	508	121	WGS
13	陈连勇等^[23]	2016	846	146	PCR扩增
14	赵刚等^[24]	2016—2019	3317	197	PCR扩增
15	文书等^[25]	2015—2017	78	55	PCR扩增
16	许河南等^[26]	2019—2020	242	57	荧光PCR熔解曲线
17	蒋燕成等^[27]	2019—2021	51	50	DNA微阵列芯片法
18	穆成等^[28]	2020	72	72	WGS
19	邓丽等^[29]	2022	380	246	PCR扩增
20	Islam等^[30]	2015—2017	206	191	PCR扩增
21	Zhdanova等^[31]	2023	137	80	WGS
22	Jia等^[32]	2007—2016	913	264	PCR扩增
总计	-	-	12923	2824	-

地区	菌株数 (株)	构成比 (%)	耐INH (株)	构成比 (%)	katG基因^a (株)	构成比 (%)	inhA基因^b (株)	构成比 (%)
华北	90	0.69	31	1.08	29	1.35	2	0.46
华东	7755	60.00	1400	49.58	847	40.22	230	53.36
华中	277	2.14	156	5.53	121	5.75	25	5.80
华南	1996	15.45	623	22.06	401	19.04	113	26.22
西南	2805	21.71	614	21.75	708	33.62	61	14.16
合计	12923	100.00	2824	100.00	2106	100.00	431	100.00

地区	菌株数	构成比 (%)	耐INH (株)	构成比 (%)	katG基因^a (株)	构成比 (%)	inhA基因^b (株)	构成比 (%)
东部	9533	73.76	2004	70.96	1241	58.90	343	79.58
中部	585	4.53	206	7.29	162	7.72	26	6.03
西部	2805	21.71	614	21.75	703	33.38	62	14.39
合计	12923	100.00	2824	100.00	2106	100.00	431	100.00

耐药检测结果	菌株数(株)	构成比(%)
产生耐药类型
野生型^a	153	5.42(153/2824)
突变型	2638	93.41(2638/2824)
其他^b	33	1.17(33/2824)
异烟肼耐药类型
异烟肼单耐药	759	26.88(759/2824)
非异烟肼耐药	2065	73.12(2065/2824)
表型药敏与分子药敏结果相同		87.76(11251/12923)
表型耐药+分子耐药	2638
表型敏感+分子敏感	8613
表型药敏与分子药敏结果不同		5.20(627/12923)
表型耐药+分子敏感	186
表型敏感+分子耐药	441
其他^c	1045	8.09(1045/12923)