中国西北五省结核分枝杆菌耐药基因突变特征分析

doi:10.19982/j.issn.1000-6621.20250458

中国防痨杂志 ›› 2026, Vol. 48 ›› Issue (6): 856-864.doi: 10.19982/j.issn.1000-6621.20250458

中国西北五省结核分枝杆菌耐药基因突变特征分析

蒋学峰¹, 周临军², 孙焱³, 蒋明霞⁴, 马玲⁵, 梅蠡⁶, 刘东鑫⁷()

¹ 宁夏回族自治区第四人民医院, 银川 750021
² 新疆维吾尔自治区喀什地区肺科医院, 喀什 844000
³ 陕西省疾病预防控制中心结核病麻风病预防控制所, 西安 710054
⁴ 青海省疾病预防控制中心结核病预防控制所, 西宁 810000
⁵ 甘肃省疾病预防控制中心结核病预防控制所, 兰州 730000
⁶ 北京市疾病预防控制中心结核病预防控制所, 北京 100035
⁷ 中国疾病预防控制中心实验室管理处, 北京 102206

收稿日期:2025-11-24 出版日期:2026-06-10 发布日期:2026-05-25
通信作者: 刘东鑫 E-mail:liucommon@126.com
基金资助:
宁夏自然科学基金项目(2024AAC05094);甘肃省自然科学基金项目(22JR5RA712)

Analysis of drug resistance gene mutation characteristics in Mycobacterium tuberculosis strains from five northwestern provinces of China

Jiang Xuefeng¹, Zhou Linjun², Sun Yan³, Jiang Mingxia⁴, Ma Ling⁵, Mei Li⁶, Liu Dongxin⁷()

¹ The Fourth People’s Hospital of Ningxia Hui Autonomous Region, Yinchuan 750021, China
² Pulmonary Hospital of Kashi Region, Xinjiang Uygur Autonomous Region, Kashgar 844000, China
³ Tuberculosis and Leprosy Prevention and Control Institute, Shaanxi Center for Disease Control and Prevention, Xi’an 710054, China
⁴ Tuberculosis Prevention and Control Institute, Qinghai Provincial Center for Disease Control and Prevention, Xining 810000, China
⁵ Tuberculosis Prevention and Control Institute, Gansu Provincial Center for Disease Control and Prevention, Lanzhou 730000, China
⁶ Tuberculosis Prevention and Control Institute, Beijing Municipal Center for Disease Control and Prevention, Beijing 100035, China
⁷ Laboratory Management Office, Chinese Center for Disease Control and Prevention, Beijing 102206, China

Received:2025-11-24 Online:2026-06-10 Published:2026-05-25
Contact: Liu Dongxin E-mail:liucommon@126.com
Supported by:
Natural Science Foundation of Ningxia(2024AAC05094);Natural Science Foundation of Gansu Province(22JR5RA712)

摘要/Abstract

摘要：

目的: 分析中国西北五省结核分枝杆菌(Mycobacterium tuberculosis,MTB)的耐药相关基因突变特征,评估全基因组测序(whole genome sequencing,WGS)技术预测表型耐药性的效能,探讨现有快速分子诊断方法在该地区的适用性,为区域耐药结核病防控提供科学依据。方法: 收集2013—2022年间来自陕西省、甘肃省、宁夏回族自治区、青海省、新疆维吾尔自治区的2125株MTB临床分离株,采用微孔板法对利福平、异烟肼等13种抗结核药物进行表型药物敏感性试验(简称“表型药敏试验”),同时进行WGS与耐药基因突变分析。以表型药敏结果为金标准,分析WGS预测表型耐药的敏感度和特异度,分析区域耐药突变特征谱与快速分子诊断方法未覆盖位点。结果: 在1716株获得表型药敏结果的菌株中,利福平和异烟肼的耐药占比分别为5.30%(91/1716)和10.72%(184/1716)。WGS预测利福平、异烟肼、乙胺丁醇、阿米卡星、卡那霉素、左氧氟沙星、莫西沙星、乙硫异烟胺、贝达喹啉、氯法齐明和链霉素的敏感度分别为96.74%(89/92)、72.83%(134/184)、67.92%(36/53)、5/6、7/8、54.84%(17/31)、59.09%(13/22)、80.00%(24/30)、1/8、23.08%(3/13)和97.22%(35/36),特异度均高于98.34%。西北地区利福平和异烟肼最主要的耐药突变分别是rpoB_Ser450Leu(58.16%,57/98)和katG_Ser315Thr(68.71%,101/147),但同样存在rpoB_Ser450Trp、katG_Ser315Ile、katG_Pro429fs、rpoB_His445_Lys446delinsGln等一些稀有突变。快速分子检测方法对利福平、异烟肼、氟喹诺酮类药物耐药位点的覆盖率分别为97.94%(95/97)、93.01%(133/143)和78.95%(15/19)。结论: WGS对中国西北地区MTB传统抗结核药物耐药性的预测价值较高,该地区存在较多稀有突变位点,提示需进一步扩展分子诊断方法的靶点覆盖范围,以提升耐药分子诊断的准确率,从而阻断耐药结核病的传播。

关键词: 分枝杆菌, 结核, 抗药性, 基因, 突变

Abstract:

Objective: To analyze the characteristics of drug resistance-related gene mutations in Mycobacterium tuberculosis (MTB) isolated from five provinces in Northwest China, evaluate the efficacy of whole genome sequencing (WGS) technology in predicting phenotypic drug resistance, explore the applicability of existing rapid molecular diagnostic methods in this region, and provide scientific basis for the prevention and control of regional drug-resistant tuberculosis. Methods: From 2013 to 2022, 2125 clinical isolates of MTB were collected from Shaanxi Province, Gansu Province, Ningxia Hui Autonomous Region, Qinghai Province, and Xinjiang Uygur Autonomous Region. Phenotypic drug susceptibility testing was performed using the microplate method for 13 anti-tuberculosis drugs. Simultaneously, WGS and drug resistance gene mutation analysis were conducted. Taking the phenotypic drug susceptibility results as the gold standard, the sensitivity and specificity of WGS in predicting phenotypic drug resistance were analyzed. Additionally, the regional drug resistance mutation spectrum and sites not covered by rapid molecular diagnostic methods were analyzed. Results: Among the 1716 strains with phenotypic drug susceptibility results, the resistance rates of rifampin and isoniazid were 5.30% (91/1716) and 10.72% (184/1716), respectively. WGS predicted the sensitivity of rifampin, isoniazid, ethambutol, amikacin, kanamycin, levofloxacin, moxifloxacin, ethionamide, bedaquiline, clofazimine, and streptomycin to be 96.74% (89/92), 72.83% (134/184), 67.92% (36/53), 5/6, 7/8, 54.84% (17/31), 59.09% (13/22), 80.00% (24/30), 1/8, 23.08% (3/13), and 97.22% (35/36), respectively, with specificities all exceeding 98.34%. The most prevalent resistance mutations to rifampin and isoniazid in the northwest region were rpoB_Ser450Leu (58.16%, 57/98) and katG_Ser315Thr (68.71%, 101/147), respectively, but there were also some rare mutations such as rpoB_Ser450Trp, katG_Ser315Ile, katG_Pro429fs, rpoB_His445_Lys446delinsGln. The coverage rates of rapid molecular detection methods for resistance loci of rifampin, isoniazid, and fluoroquinolones were 97.94% (95/97), 93.01% (133/143), and 78.95% (15/19), respectively. Conclusion: WGS has a high predictive value for the resistance of MTB to traditional anti-tuberculosis drugs in Northwest China. The presence of numerous rare mutation sites in this region suggests the need to further expand the target coverage of molecular diagnostic methods to improve the accuracy of drug-resistant molecular diagnosis, thereby blocking the transmission of drug-resistant tuberculosis.

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

中图分类号:

蒋学峰, 周临军, 孙焱, 蒋明霞, 马玲, 梅蠡, 刘东鑫. 中国西北五省结核分枝杆菌耐药基因突变特征分析[J]. 中国防痨杂志, 2026, 48(6): 856-864. doi: 10.19982/j.issn.1000-6621.20250458

Jiang Xuefeng, Zhou Linjun, Sun Yan, Jiang Mingxia, Ma Ling, Mei Li, Liu Dongxin. Analysis of drug resistance gene mutation characteristics in Mycobacterium tuberculosis strains from five northwestern provinces of China[J]. Chinese Journal of Antituberculosis, 2026, 48(6): 856-864. doi: 10.19982/j.issn.1000-6621.20250458

图/表 5

表1

表2

表3

表4

我国西北五省结核分枝杆菌耐药位点组成情况

药品/位点	突变株数(突变率,%)
利福平
rpoB_Asp435Tyr	3(3.06)
rpoB_Asp435Val	1(1.02)
rpoB_Gln432Leu	1(1.02)
rpoB_His445_Lys446delinsGln	1(1.02)
rpoB_His445Arg	1(1.02)
rpoB_His445Asn	3(3.06)
rpoB_His445Asn、rpoB_Leu452Val	1(1.02)
rpoB_His445Asp	6(6.12)
rpoB_His445Leu	1(1.02)
rpoB_His445Tyr	5(5.10)
rpoB_Ile491Phe	1(1.02)
rpoB_Leu430Pro	5(5.10)
rpoB_Leu430Pro、rpoB_Asp435Ala	1(1.02)
rpoB_Leu430Pro、rpoB_Asp435Tyr	1(1.02)
rpoB_Leu430Pro、rpoB_Met434Ile	1(1.02)
rpoB_Leu443Trp、rpoB_Thr444Pro、 rpoB_His445_Lys446delinsGln	1(1.02)
rpoB_Leu452Pro	5(5.10)
rpoB_Ser441Gln	1(1.02)
rpoB_Ser450Leu	57(58.16)
rpoB_Ser450Trp	1(1.02)
rpoB_Val170Phe	1(1.02)
合计	98(100.00)
异烟肼
inhA_-154G>A、katG_Ser315Thr	1(0.68)
inhA_-77-T>A	1(0.68)
inhA_-77-T>C	2(1.36)
inhA_-777C>T	23(15.65)
inhA_-777C>T、katG_Ser315Thr	1(0.68)
inhA_-779G>T	1(0.68)
katG_His25fs	3(2.04)
katG_Pro429fs	1(0.68)
katG_Ser315Arg	1(0.68)
katG_Ser315Asn	8(5.44)
katG_Ser315Ile	1(0.68)
katG_Ser315Thr	101(68.71)
katG_Ser94Ala	1(0.68)
katG_Trp204*	1(0.68)
katG_Val22fs	1(0.68)
合计	147(100.00)
乙胺丁醇
embB_Asp354Ala	1(2.63)
embB_Gln497Arg	3(7.89)
embB_Gly406Ala	8(21.05)
embB_Gly406Ser	1(2.63)
embB_Met306Ile	16(42.11)
embB_Met306Ile、embB_Gly406Ser	1(2.63)
embB_Met306Val	8(21.05)
合计	38(100.00)
氟喹诺酮类药物
gyrB_Ala504Val	1(4.76)
gyrA_Ala90Val	5(23.81)
gyrB_Asp461Asn	2(9.52)
gyrB_Asp461His	1(4.76)
gyrA_Asp94Ala	1(4.76)
gyrA_Asp94Asn	3(14.29)
gyrA_Asp94Gly	2(9.52)
gyrA_Asp94Tyr	2(9.52)
gyrB_Ser447Phe	1(4.76)
gyrA_Ser91Pro	2(9.52)
gyrA_Ala90Val、gyrA_Asp94Gly	1(4.76)
合计	21(100.00)
阿米卡星
rrs_1401A>G	5(5/5)
卡那霉素
eis_-10G>A	2(2/7)
rrs_1401A>G	5(5/7)
合计	7(7/7)
乙硫异烟胺
inhA_-154G>A、ethA_His281fs	1(2.63)
inhA_-770T>A	1(2.63)
inhA_-770T>C	2(5.26)
inhA_-777C>T	24(63.16)
inhA_-779G>T	1(2.63)
ethA_Gln206*	1(2.63)
ethA_Gln271*	1(2.63)
ethA_His119fs	1(2.63)
ethA_Leu82fs	3(7.89)
ethA_Ser94Ala	1(2.63)
ethA_Trp240*	1(2.63)
ethA_Tyr382*	1(2.63)
合计	38(100.00)

表4

表5

参考文献 19

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方法	药品	基因	位点范围
MeltPro TB	异烟肼	katG	315密码子
MeltPro TB	异烟肼	inhA启动子	-17~-8位核苷酸
MeltPro TB	异烟肼	ahpC	-44~-30及-15~3位核苷酸
MeltPro TB	利福平	rpoB	507~533位氨基酸密码子
MeltPro TB	氟喹诺酮类	gyrA	87~95密码子
MeltPro TB	氟喹诺酮类	gyrB	492~505密码子
MeltPro TB	阿米卡星	rrs	1396~1417位核苷酸
MeltPro TB	卡那霉素	rrs	1396~1417位核苷酸
MeltPro TB	卡那霉素	eis启动子	-6~-42位核苷酸

药品	表型耐药 (株)		表型敏感 (株)		合计 (株)		敏感度 (%,95%CI)		特异度 (%,95%CI)
利福平							96.74(90.09~99.15)		99.50(98.99~99.77)
基因型耐药	89		8		97
基因型敏感	3		1616		1619
合计	92		1624		1716
异烟肼							72.83(65.69~78.98)		99.41(98.84~99.71)
基因型耐药	134		9		143
基因型敏感	50		1523		1573
合计	184		1532		1716
乙胺丁醇							67.92(53.55~79.70)		100.00(99.71~100.00)
基因型耐药	36		0		36
基因型敏感	17		1663		1680
合计	53		1663		1716
阿米卡星							5/6		99.65(99.72~100.00)
基因型耐药	5		0		5
基因型敏感	1		1710		1711
合计	6		1710		1716
卡那霉素							7/8		99.53(99.72~100.00)
基因型耐药	7		0		7
基因型敏感	1		1708		1709
合计	8		1708		1716
莫西沙星							59.09(36.68~78.52)		99.65(99.19~99.86)
基因型耐药	13		6		19
基因型敏感	9		1688		1697
合计	22		1694		1716
左氧氟沙星								54.84(36.30~72.22)		99.88(99.52~99.98)
基因型耐药		17		2		19
基因型敏感		14		1683		1697
合计		31		1685		1716
贝达喹啉								1/8		98.34(0.66~53.32)
基因型耐药		1		26		27
基因型敏感		7		1536		1543
合计		8		1562		1570
氯法齐明								23.08(6.16~54.02)		99.87(99.48~99.98)
基因型耐药		3		2		5
基因型敏感		10		1555		1565
合计		13		1557		1570
德拉马尼								2.86(0.15~16.62)		99.96(99.83~99.99)
基因型耐药		1		2		3
基因型敏感		34		4661		4695
合计		35		4663		4698
乙硫异烟胺								80.00(60.87~91.60)		99.17(98.57~99.53)
基因型耐药		24		14		38
基因型敏感		6		1672		1678
合计		30		1686		1716
链霉素								97.22(83.80~99.85)		98.53(94.25~99.74)
基因型耐药		35		2		37
基因型敏感		1		134		135
合计		36		136		172

中国西北五省结核分枝杆菌耐药基因突变特征分析

Analysis of drug resistance gene mutation characteristics in Mycobacterium tuberculosis strains from five northwestern provinces of China

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药品	流行病学界值(mg/L)
异烟肼	0.10
利福平	0.50
乙胺丁醇	4.00
莫西沙星	1.00
左氧氟沙星	1.00
卡那霉素	4.00
阿米卡星	1.00
乙硫异烟胺	4.00
氯法齐明	0.25
利奈唑胺	1.00
德拉马尼	0.12
贝达喹啉	0.25
链霉素	2.00

药品	第1组^a	第2组^a	多突变	合计
利福平
株数	102	2	6	110
突变率(%)	92.73	1.82	5.46
异烟肼
株数	158	13	2	173
突变率(%)	91.33	7.51	1.16
乙胺丁醇
株数	158	0	1	159
突变率(%)	91.33	0.00	2.22
左氧氟沙星
株数	19	3	0	22
突变率(%)	86.36	13.64	0.00
莫西沙星
株数	17	5	0	22
突变率(%)	77.27	22.73	0.00
贝达喹啉
株数	4	29	0	33
突变率(%)	12.12	87.88	0.00
氯法齐明
株数	1	32	0	33
突变率(%)	3.03	96.97	0.00
阿米卡星
株数	7	0	0	7
突变率(%)	100.00	0.00	0.00
卡那霉素
株数	9	0	0	9
突变率(%)	100.00	0.00	0.00
链霉素
株数	177	20	2	199
突变率(%)	88.95	10.05	1.01
乙硫异烟胺
株数	29	9	5	43
突变率(%)	67.44	20.93	11.63
吡嗪酰胺
株数	23	5	0	28
突变率(%)	82.14	17.86	0.00

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[4]	张姗姗, 韩亚轩, 张小燕, 李建英, 任斐, 张耀辉, 杨海霞. 肺癌共病耐药肺结核患者的临床特征及生存分析[J]. 中国防痨杂志, 2026, 48(5): 641-650.
[5]	李芹, 李林宏, 刘娇, 冯美, 谢祺, 张桂仙, 杨琼, 杨丹丹. AIDS合并利福平耐药结核性乳腺脓肿一例及文献复习[J]. 中国防痨杂志, 2026, 48(5): 687-693.
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