Study on high-concentration p-aminosalicylic acid resistant Mycobacterium tuberculosis induced in vitro and the mutation sites

doi:10.19982/j.issn.1000-6621.20220457

Abstract

Abstract:

Objective: To explore the potential mechanisms of action and resistance to para-aminosalicylic acid (PAS) by sequencing the whole genome (WGS) of lab-induced PAS-resistant Mycobacterium tuberculosis (MTB) and clinical isolates. Methods: The MTB standard strain was induced into standard PAS resistant strain and high-level drug resistant strain by in vitro drug concentration gradient induction method, and the induced strains of each generation were collected and preserved. The minimum inhibitory concentration (MIC) and cross resistance of the above induced strains were detected by liquid microplate method, and clinical isolates with similar drug resistance levels were screened. By sequencing the whole genome of PAS resistant laboratory mutants and clinical isolates, the mechanism of resistance of MTB to PAS was explored at the gene level. Results: A dynamic PAS-resistant model of MTB was established in vitro. The WGS results showed that PAS resistance may be directly related to three site mutations, plcC (Q462R), folC (S150R) and a thyA upstream site (3074495G→A). Except for genetic mutations, there were other regulatory mechanisms for the resistance to high levels PAS of MTB. Moreover, WGS results of PAS-susceptible and resistant clinical isolates revealed that some folC, thyA and thyX mutations were not caused by PAS. Conclusion: The PAS-resistant model of MTB constructed in this study provides an ideal biological model for the research and clinical practice of PAS-resistant tuberculosis. WGS analysis has clarified the resistance mechanism of MTB to PAS at the DNA level, and other regulatory mechanisms still need further investigation. The gene mutations associated with PAS resistance obtained in this study also need further experimental validation.

Key words: Mycobacterium tuberculosis, Para-aminosalicylic acid, Drug resistance, bacterial, Genes, bacterial, Mutation

CLC Number:

R52
R453

Yu Meiling, Zhang Chenchen, Wei Wenjing, Zhao Yuchuan, Zhuo Wenji, Zheng Lei. Study on high-concentration p-aminosalicylic acid resistant Mycobacterium tuberculosis induced in vitro and the mutation sites[J]. Chinese Journal of Antituberculosis, 2023, 45(1): 60-66. doi: 10.19982/j.issn.1000-6621.20220457

Figures/Tables 3

菌株	INH (MIC= 0.2)	Sm (MIC= 2)	EMB (MIC= 5)	Ofx (MIC= 2)	Mfx (MIC= 0.5)	Am (MIC= 1)	Km (MIC= 5)	Cm (MIC= 2)	Pto (MIC= 2.5)	PAS (MIC= 1)	RFP (MIC= 1)	Rfb (MIC= 0.5)	Lfx (MIC= 2)	PZA (MIC= 100)
P1	<0.025	0.50	1.25	0.5	0.12	1	2.50	1	<0.62	<0.5	<0.25	<0.12	<0.5	<50
P2	<0.025	0.50	1.25	1.0	0.12	1	2.50	1	<0.62	<0.5	<0.25	<0.12	<0.5	<50
P3	<0.025	<0.25	1.25	1.0	0.25	1	2.50	2	<0.62	4.0	0.50	<0.12	<0.5	<50
P4	0.200	0.50	1.25	1.0	0.12	1	1.25	1	<0.62	4.0	0.50	<0.12	<0.5	<50
P5	0.200	0.50	1.25	1.0	0.12	1	2.50	1	<0.62	4.0	0.50	<0.12	<0.5	<50
P6	<0.025	<0.25	1.25	1.0	0.12	1	2.50	2	<0.62	8.0	<0.25	<0.12	<0.5	<50
P7	<0.025	<0.25	1.25	1.0	0.25	1	2.50	2	<0.62	8.0	<0.25	<0.12	<0.5	<50
P8	<0.025	<0.25	1.25	1.0	0.12	1	2.50	2	<0.62	16.0	<0.25	<0.12	<0.5	<50
P9	<0.025	0.50	1.25	1.0	0.12	1	2.50	2	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P10	<0.025	<0.25	1.25	1.0	0.12	1	2.50	1	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P11	<0.025	<0.25	1.25	1.0	0.12	1	2.50	1	1.25	>16.0	<0.25	<0.12	<0.5	<50
P12	0.100	0.50	2.50	1.0	0.25	1	5.00	2	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P13	<0.025	<0.25	1.25	1.0	0.12	1	2.50	2	<0.62	16.0	0.50	<0.12	<0.5	<50
P14	0.050	0.50	1.25	1.0	0.12	1	2.50	2	<0.62	16.0	0.50	<0.12	<0.5	<50
P15	<0.025	0.50	1.25	1.0	0.12	1	2.50	2	<0.62	16.0	<0.25	<0.12	<0.5	<50
P16	<0.025	0.50	1.25	1.0	0.12	1	2.50	2	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P17	<0.025	0.50	1.25	1.0	0.12	1	2.50	2	1.25	>16.0	0.50	<0.12	<0.5	<50
P18	<0.025	0.50	1.25	1.0	0.12	1	2.50	2	<0.62	16.0	<0.25	<0.12	<0.5	<50
P19	0.050	<0.25	1.25	1.0	0.12	1	2.50	1	<0.62	16.0	<0.25	<0.12	<0.5	<50
P20	<0.025	<0.25	1.25	1.0	0.12	1	2.50	1	<0.62	16.0	<0.25	<0.12	<0.5	<50
P21	<0.025	<0.25	1.25	0.5	0.12	1	2.50	1	<0.62	16.0	<0.25	<0.12	<0.5	<50
P22	0.050	<0.25	1.25	1.0	0.12	1	2.50	2	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P23	0.050	0.50	1.25	1.0	0.12	1	2.50	2	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P24	<0.025	<0.25	1.25	1.0	0.12	1	2.50	1	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P25	<0.025	0.50	1.25	1.0	0.12	1	2.50	1	<0.62	>16.0	<0.25	<0.12	<0.5	<50
P26	0.050	<0.25	1.25	1.0	0.25	1	2.50	1	<0.62	>16.0	<0.25	<0.12	<0.5	<50

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