Molecular characterization and risk factors of cross-resistance between isoniazid and prothionamide in multidrug-resistant Mycobacterium tuberculosis in Ningbo, China

doi:10.19982/j.issn.1000-6621.20250414

Abstract

Abstract:

Objective: This study aimed to investigate the characterization, the drug resistance-associated gene mutations in multidrug-resistant Mycobacterium tuberculosis (MDR-MTB) strains based on whole-genome sequencing (WGS) and to analyze the risk factors of cross-resistance between isoniazid (INH) and prothionamide (Pto). Methods: A total of 134 MDR-MTB isolates selected in the drug resistance survey in Ningbo between 2021 and 2024 were enrolled in the study. WGS was conducted to detect the genomic DNA extracted from clinical isolates. The sequencing data were analyzed using an in-house pipeline and the gene mutations determined based on MTB drug resistance gene database. The conventional drug susceptibility test was also used to detect the drug-resistant profiles against 9 antibiotics (levofloxacin, bedaquiline, linezolid, delamanid, clofazimine, amikacin, capreomycin, para-aminosalicylic acid and Pto). Combined with phenotypic drug susceptibility testing results and clinical data, the cross-resistance profile and risk factors between INH and Pto among MDR-MTB isolates were comprehensively analyzed. Results: Among 134 MDR-MTB isolates, 129 (96.3%, 129/134) carried INH resistance-associated mutations. Mutations were present in all three INH resistance-associated genes, katG, inhA and ahpC. The main mutation types were katG Ser315Thr (89.1%, 98/110), inhA c-15t (50.0%, 17/34) and ahpC c-81t (4/9). Among 134 MDR-MTB isolates, the overall Pto resistance rate (INH-Pto cross-resistance rate) was 20.2% (27/134), with 92.6% (25/27) harboring mutations. The gene mutations associated with Pto resistance were ethA and inhA, with mutation rates of 63.0% (17/27) and 77.8% (21/27), respectively. The major mutation types were ethA 11_12insA (70.6%, 12/17) and inhA t-8C (66.7%, 14/21). Among MDR-MTB and Pre-XDR-MTB, the INH-Pto cross-resistance rates were 20.2% (27/134) and 43.6% (24/55). Multivariate logistic regression showed that moxifloxacin given (OR (95%CI): 4.123 (1.207-14.080)) and the resistance of levofloxacin (OR (95%CI): 0.075 (0.019-0.293)) were risk factors for INH-Pto cross-resistance. Conclusion: The prevalence of INH-Pto cross-resistance among MDR-MTB was serious in Ningbo and the mutation type inhA t-8C was closely with INH-Pto cross-resistance. The regimen for INH-Pto cross-resistant patients should be according to the drug susceptibility results of fluoroquinolones and choose optimal treatment.

Key words: Mycobacterium tuberculosis, Drug resistance, multiple, bacterial, Prothionamide, Mutation, Genes

CLC Number:

Che Yang, Zhang Dongliang, Li Xiangchen, Lu Yewei, Liu Zhengwei. Molecular characterization and risk factors of cross-resistance between isoniazid and prothionamide in multidrug-resistant Mycobacterium tuberculosis in Ningbo, China[J]. Chinese Journal of Antituberculosis, 2026, 48(5): 576-585. doi: 10.19982/j.issn.1000-6621.20250414

Figures/Tables 5

特征		非交叉耐药组(107例)			交叉耐药组(27例)			χ²值			P值
性别								5.048			0.025
男性		77(72.0)			25(92.6)
女性		30(28.0)			2(7.4)
年龄组(岁)								3.679			0.159
15~29		21(19.6)			10(37.0)
30~59		60(56.1)			12(44.4)
≥60		26(24.3)			5(18.6)
户籍								0.091			0.763
本地		52(48.6)			14(51.9)
非本地		55(51.4)			13(48.1)
吸烟								0.518			0.472
是		21(19.6)			7(25.9)
否		86(80.4)			20(74.1)
饮酒									0.790			0.374
是			14(13.1)			6(22.2)
否			93(86.9)			21(77.8)
合并糖尿病									0.036			0.850
是			22(20.6)			6(22.2)
否			85(79.4)			21(77.8)
合并高血压									0.076			0.783
是			14(13.1)			3(11.1)
否			93(86.9)			24(88.9)
合并慢性阻塞性肺疾病									0.313			0.576
是			3(2.8)			2(7.4)
否			104(97.2)			25(92.6)
结核病治疗史									3.452			0.063
有			46(43.0)			17(63.0)
无			61(57.0)			10(37.0)
空洞									1.570			0.210
有			57(53.3)			18(66.7)
无			50(46.7)			9(33.3)
涂片镜检									0.223			0.636
阳性			53(49.5)			12(44.4)
阴性			54(50.5)			15(55.6)
诊断时生化指标
A/G									3.622			0.057
正常			69(64.5)			12(44.4)
异常			38(35.5)			15(55.6)
AST/ALT									0.017			0.895
正常			50(46.7)			13(48.1)
异常			57(53.3)			14(51.9)
CRP水平									0.831			0.362
正常			61(57.0)			18(66.7)
异常			46(43.0)			9(33.3)
CRE水平									2.720			0.099
正常			65(60.7)			21(77.8)
异常			42(39.3)			6(22.2)
BUN水平									1.269			0.260
正常			85(79.4)			24(88.9)
异常			22(20.6)			3(11.1)
TC水平									1.014			0.314
正常			100(93.5)			23(85.2)
异常			7(6.5)			4(14.8)
TG水平								0.158			0.691
正常		100(93.5)			24(88.9)
异常		7(6.5)			3(11.1)
用药情况
使用Bdq								0.347			0.556
是		34(31.8)			7(25.9)
否		73(68.2)			20(74.1)
使用Lzd								0.019			0.892
是		61(57.0)			15(55.6)
否		46(43.0)			12(44.4)
使用Mfx								4.497			0.034
是		52(48.6)			7(25.9)
否		55(51.4)			20(74.1)
使用Lfx								0.016			0.900
是		25(23.4)			6(22.2)
否		82(76.6)			21(77.8)
使用Cfz								0.094			0.759
是		48(44.9)			13(48.1)
否		59(55.1)			14(51.9)
使用Cm								0.008			0.931
是		5(4.7)			2(7.4)
否		102(95.3)			25(92.6)
使用Cs								0.005			0.946
是		76(71.0)			19(70.4)
否		31(29.0)			8(29.6)
使用PAS								5.343			0.021
是		4(3.7)			5(18.5)
否		103(96.3)			22(81.5)
使用Pto								0.078			0.780
是		25(23.4)			7(25.9)
否		82(76.6)			20(74.1)
使用PZA								0.461			0.497
是		72(67.3)			20(74.1)
否		35(32.7)			7(25.9)
菌株基因型								2.969			0.085
Lineage 2		92(86.0)			27(100.0)
Lineage 4		15(14.0)			0(0.0)
菌株耐药谱
耐Lfx								31.986			<0.010
是		31(29.0)			24(88.9)
否		76(71.0)			3(11.1)
耐Am							17.184			<0.010
是	3(2.8)			8(29.6)
否	104(97.2)			19(70.4)
耐Cm							5.343			0.021
是	4(3.7)			5(18.5)
否	103(96.3)			22(81.5)
耐PAS							0.420			0.517
是	20(18.7)			3(11.1)
否	87(81.3)			24(88.9)

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因素	β值	s${}_{\overline{x}}$值	Wald χ²值	P值	OR(95%CI)值
性别	-1.389	0.889	2.439	0.118	0.249(0.044~1.425)
使用Mfx	1.416	0.627	5.109	0.024	4.123(1.207~14.080)
使用PAS	-0.694	1.002	0.480	0.489	0.499(0.070~3.562)
耐Lfx	-2.588	0.693	13.929	<0.010	0.075(0.019~0.293)
耐Am	-2.420	1.430	2.862	0.091	0.089(0.005~1.467)
耐Cm	0.249	1.669	0.022	0.882	1.282(0.049~33.799)