Epidemiology of Nontuberculous Mycobacteria in Nanjing and MAB_0540 Mutations Associated with Clofazimine Resistance in Mycobacterium abscessus

Figures & data

Figure 1 Distribution of NTM species.

Figure 2 The changes of six major NTM strains from 2019 to 2021. The number of patients infected with each strain of NTM annually from 2019 to 2021 was shown in (A–C). The ratio of males to females in each strain of NTM for each year was displayed (a–c).

Abbreviations: MIN, M. intracellulare; MAV, M. avium; MAB, M. abscessus; MKA, M. kansasii; MFO, M. fortuitum/M. suis; MGO, M. gordonae.

Table 1 Gender and Age of Patients Infected with NTM

Age Group	Male	Female	Total	χ^2	P
<20	3	4	7 (1.27%)	0.143	0.705
20–29	22	16	38 (6.91%)	0.947	0.330
30–39	47	20	67 (12.18%)	10.881	0.001
40–49	32	17	49 (8.91%)	4.592	0.032
50–59	48	63	111 (20.18%)	2.027	0.155
60–69	67	75	142 (25.82%)	0.451	0.502
70–79	52	45	97 (17.64%)	0.505	0.477
≥80	27	12	39 (7.09%)	5.769	0.016
Total	298	252	550 (100%)	21.619	0.003
Past medical history and complications		Number of people		Proportion
Tuberculosis (TB)		256		46.55%
Nontuberculous mycobacteriosis		321		58.36%
Immune deficiency disease		73		13.27%
Bronchiectasis		43		7.82%
Hepatic dysfunction		205		37.27%
Diabetes		20		3.64%
Hypertension		12		2.18%
Tumor + surgical history		22		4.00%
Coronary heart disease (CHD)		7		1.27%
Uremia		2		0.36%
Leucopenia		31		5.64%
Cytomegalovirus infection		8		1.45%

Table 2 Gender Distribution Among Different Species Types

Species Type	Gender		χ^2	P
	Male	Female
M. avium	71	32	14.767	0.000
M. kansasii	33	3	25.000	0.000
M. intracellulare	143	158	0.748	0.387
M. abscessus	28	49	5.727	0.017
M. fortuitum/porcine	14	9	1.087	0.297
M. gordonae	25	18	1.140	0.286
M. szulgai	4	1	---	---
M. triviale	1	0	---	---
M. scrofulaceum	2	1	---	---
M. xenia	1	4	---	---

Figure 3 MICs distributions of M. abscessus and M. intracellulare to bedaquiline, clofazimine, linezolid, delamanid and pretomanid. (A) The MIC distributions of M. abscessus against bedaquiline. (B) The MIC distributions of M. intracellulare against bedaquiline. (C) The MIC distributions of M. abscessus against clofazimine. (D) The MIC distributions of M. intracellulare against clofazimine. (E) The MIC distributions of M. abscessus against linezolid. (F) The MIC distributions of M. intracellulare against linezolid. (G) The MIC distributions of M. abscessus against delamanid. (H) The MIC distributions of M. intracellulare against delamanid. (I) The MIC distributions of NTM against pretomanid. The position indicated by the arrow indicates the critical concentration value of resistance.

Abbreviations: MAB, M. abscessus; MIN, M. intracellulare; MFO, M. fortuitum/porcine; MGO, M. gordonii; BDQ, bedaquiline; CFZ, clofazimine; LZD, linezolid; DLM, delamanid; PA-824, pretomanid.

Table 3 NTM Mutation M. abscessus of BDQ, CFZ, and LZD

Drug	Gene	Strains Numbers	Nucleotid Mutation	Amino Acid Change
BDQ	MAB_1448	1	13insertC	Frameshift mutation
		2	WT	WT
	MAB_4384	1	A180G, C264T, A549G	WT
		2	Failure of amplification	--
	MAB_2299c	2	A264C, T612G	WT
		1	A264C, T612G, A15C, A57G, A99G, G165C, G270C, C300G, C303T, G309C, A339G, C345T, T366C, A513G, C573G, G630A, A225G	WT
CFZ	MAB_2299c	2	WT	WT
		1	A264C, T612G, A15C, A57G, A99G, G165C, G270C, C300G, C303T, G309C, A339G, C345T, T366C, A513G, C573G, G630A, A225G	WT
		1	A264C	WT
		4	A264C, T612G	WT
	MAB_1483	2	T183C, G228C, A243G, C252T, T267C, A282G, C378T, A435G, C483T, T489C, C501T, C576T, C636T, T687C, G735A	WT
		7	WT	WT
		1	G285A	WT
		1	T183C, G228C, A282G, C288T, C342T	WT
	MAB_0540	2	C29G, T105C, T219C, C387A, G391A, C687T, G771C, G909A, A135G, C369T	Missense mutations
		2	WT	WT
		1	C29G, T105C, T219C, C387A, G391A, C687T, G771C, G909A	Missense mutations
		2	T105C, T219C, G391A, C687T, G771C, A135G, 30–41 deletion, A42G, C69A, G78A, C96A, C133A, A151G, T159G, A261G, G316T, A385G, G391A, C412T, G429A, T438C, C453T, C462T, A510C, T521C, G576C, C609G, A804G, C717T, G741A, C757T, A846G, G882A, C918A, C933T, C972T, T978C	Missense mutations
		1	T105C, T219C, C387A, G391A, C389T, G771C, A135G, 30–41 deletion, C687T, G909A	Missense mutations
		3	T12G, T105C, T219C, G391A, C687T, G571T, A135G, 30–41 deletion, C795T, G909A	Missense mutations
LZD	rplC	9	WT	WT
		1	T240G	WT
	rplD	9	WT	WT
		1	T91C, C303T, G348C, T366C, G431C, T484C, C522T, G529C, T576C	Missense mutations
	rrl	9	WT	WT
		1	C634T, A1717G, T3001C	WT

Abbreviations: WT, wild type; BDQ, bedaquiline; CFZ, clofazimine; LZD, linezolid.

Table 4 NTM Mutation M. Intracellulare of LZD

Drug	NTM	Gene	Strains Numbers	Nucleotid Mutation	Amino Acid Change
LZD	M. intracellulare	rplC	19	WT	WT
			7	G360A	WT
			4	T393C	WT
			1	G372C, T393C	WT
			2	G126C, G150T, C162T, G207A, C222T, G231T, G237C, T241C, C261T, G274A, G279C, C309G, C342G, C345G, G360A, G372C, T393C, C397A, T402C, G444C, C477G, G492C, A534G, T609C, G618C	Missense mutations
		rplD	1	WT	WT
			1	C213T, C276T, G443A, A457G, G468C, T534G, C606T, A607T, G608C, T668C	Missense mutations
			2	A45G, A63C, C129G, C138G, C156T, C195G, T201C, C237G, C252G, C276T, C282T, C294T, C327G, T349C, T357C, C381T, G384A, G397A, A405G, A406C, C408G, C411G, G412T, C414G, C417T, C423G, G429C, G437A, A442C, G468C, G470A, A495G, G507A, G510C, T534G, A547G, G555T, C561T, C585G, G600C, C618G, C639G, T668C	Missense mutations
			17	T668C	WT
			8	G555A, T668C	WT
			1	G437A, T668C	WT
			4	C366G, T668C	WT

Abbreviations: WT, wild type; LZD, linezolid.

Figure 4 Gene mutations of linezolid in the 23S rRNA (rrl) among M. intracellulare clinical isolates.

Abbreviations: n, the number of mutation isolates. -, Base deletion.