Molecular Identification, and Characterization of Mycobacterium kansasii Strains Isolated from Four Tuberculosis Regional Reference Laboratories in Iran During 2016–2018

Figures & data

Table 1 Results of M. kansasii Identification by Phenotypic and Genotypic Tests

Isolates No.	Growth at 37°C	Growth on MacConkey agar	Urease production	Iron Uptake	Tween 80 Hydrolysis	Thermostable Catalase	Arylsulfatase	Niacin production	Tellurite reduction	Citrate utilization	Growth on 5% Co2	Sub-Species	hsp65-PRA BsTEII/HaeIII	Phenotypictests	Identification by rpoB
84	+	–	+	–	+	+	–	–	–	–	–	IV	235, 120, 85 /130, 115, 75, 60	M. kansasii	M. kansasii
98	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
40	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
63	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
45	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
75	+	–	– /+	–	+	+		–	–	–	–	I	235, 210/130, 105, 80	Unknown	M. kansasii
78	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
97	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
95	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
104	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
88	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
111	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
M95	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
M88	+	–	–/+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	Unknown	M. kansasii
67	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
36	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
35	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
M98	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
M63	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
33	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
19	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
21	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
32	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
34	+	–	+	–	+	+	–	–	–	–	–	I	+235, 210/130, 105, 80	M. kansasii	M. kansasii
357	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
313	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
372	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
330	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
310	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
412	+	–	+	–	+	+	–	–	–	–	–	II	235,130, 85/130,105	M. kansasii	M. kansasii
349	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
383	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii
345	+	–	+	–	+	+	–	–	–	–	–	I	235, 210/130, 105, 80	M. kansasii	M. kansasii

Table 2 Baseline Characteristics of Patients with M. kansasii-Positive Culture from Respiratory Samples

	Isolates	Sample source	A/G	PMH	Main symptoms	Genotype
1	NTM 84	BAL	60/M	HIV	Productive cough, chest wall pain & weight loss	IV
2	NTM 98	Sputum	76/F	Smoker	Productive cough	I
3	NTM 40	Sputum	57/M	Respiratory failure	Productive cough	I
4	NTM 63	Sputum	68/F	Open heart surgery	Productive cough	I
5	NTM 45	BAL	61/M	Treated tuberculosis	Productive cough	II
6	NTM 75	BAL	54/M	Normal	Local pain, small pale nodule	I
7	NTM 78	Biopsy	70/M	Pemphigus Vulgaris	Local pain, small pale nodule	I
8	NTM 97	Sputum	34/F	Renal failure	Productive cough, fever, body weight loss	I
9	NTM 95	Sputum	28/F	Normal	Fever, cough	II
10	NTM 104	Sputum	61/M	Normal	Fever	II
11	NTM 88	Sputum	60/M	Normal	Fever, inflammation and tenderness in joint	I
12	NTM 111	Sputum	70/M	COPD	Fever	I
13	MOT 95	Sputum	48/M	Normal	Productive cough, fever, weight loss	II
14	MOT 88	Sputum	81/M	Immunocompromised	Fever	II
15	NTM 67	Sputum	31/M	HIV	Fever	I
16	NTM 36	Sputum	69/M	HCV	Local abscess and discharge	I
17	NTM 35	BAL	58/M	Sarcoma	Fever, cough	II
18	MOT 98	BAL	31/M	Treated tuberculosis	Fever, cough	II
19	MOT 63	Sputum	48/M	Normal	Fever, cough	II
20	NTM 33	Sputum	71/M	Normal	Fever	I
21	NTM 19	Sputum	41/F	Normal	Local pain, small pale nodule	I
22	NTM 21	Sputum	50/M	Diabetic	Fever, cough	I
23	NTM 32	Sputum	63/F	Normal	Fever	I
24	NTM 34	Sputum	52/M	Normal	Fever	I
25	NTM 349	BAL	59/M	HIV	Fever	I
26	NTM 383	Sputum	55/M	Open heart surgery	Fever	I
27	NTM 345	Sputum	42/M	HIV	Fever, cough	I
28	NTM 357	BAL	60/M	Alcoholic	Fever	I
29	NTM 313	BAL	69/F	Treated tuberculosis	Fever	I
30	NTM 372	BAL	48/M	Open heart surgery	Fever	I
31	NTM 330	BAL	70/M	Treated tuberculosis	Fever, cough	I
32	NTM 412	BAL	89/F	Normal	Fever, cough	I
33	NTM 310	Sputum	39/M	HIV	Fever, cough	I

Abbreviations: A, age; G, gender; PMH, previous medical history; BAL, bronchoalveolar lavage; HIV, human immunodeficiency virus; HCV, hepatitis C virus.

Table 3 Frequency of M. kansasii Isolates and Subtypes by Characteristic, Symptoms at Presentation and Previous Medical History

Characteristics	M. kansasii N = 33 (%)	M. kansasii Subtypes
Year
2016	(n = 7) 21%	(I= 5) 9%,(II = 2) 6%
2017	(n = 10) 30%	(I= 7) 21%,(II = 3) 9%
2018	(n = 16) 48%	(I= 12) 36%,(II = 3) 9%, (IV = 1) 3%
Age
0–19	–	–
20–39	(n = 5) 15%	(I= 3) 9%,(II = 2) 6%
40–59	(n = 11) 33%	(I= 9)27%,(II = 3) 9%
≥60	(n = 17) 51%	(I= 12)66%, (II = 3) 9%, (IV = 1) 3%
Gender
Female	(n = 8) 24%	(I= 7)21%, (II = 1)3%
Male	(n = 25) 76%	(IV = 1) 3%,(I= 19) 57%,(II = 5) 18%
Symptoms at presentation
Fever, cough	(n = 9) 27%	(I= 5) 15 %,(II = 4) 12%
Fever	(n = 12) 36%	(I= 9) 27 %,(II = 3) 9%
Productive cough	(n = 4) 12%	(I= 3) 9 %,(II = 1) 3%
Local pain, small pale nodule	(n = 3) 9%	(I= 3) 9%
Fever, inflammation and tenderness in joint	(n = 1) 3%	(I= 1) 3%
Local abscess and discharge	(n = 1) 3%	(I= 1) 3%
Productive cough, thoracic pain & weight loss	(n =1) 3%	(IV= 1)3%
Productive cough, fever, weight loss	(n = 2) 6%	(I= 1) 3%
Underlying condition
Treated tuberculosis	(n = 4) 12%	(I= 2) 6%, (II= 2) 6%
HIV infection	(n = 5) 15%	(I= 4) 12 %, (IV= 1)3 %
Open heart surgery	(n = 3) 9%	(I= 3) 9%
COPD	(n = 1) 3%	(I= 1) 3%
HCV	(n = 1) 3%	(I= 1) 3%
Immunocompromised	(n = 1) 3%	(II=1) 3%
Renal failure	(n = 1) 3%	(I= 1) 3%
Diabetic	(n = 1) 3%	(I= 1) 3%
Pemphigus Vulgaris	(n = 1) 3%	(I= 1) 3%
Respiratory failure	(n = 1) 3%	(I= 1) 3%
Smoking	(n = 1) 3%	(I= 2) 6%
Sarcoma	(n = 1) 3%	(II= 1) 3%
Alcoholic	(n = 1) 3%	(I= 1) 3 %

Figure 1 Location and geographic distribution of M. kansasii strains during 2016–2018.

Table 4 Restriction Patterns and M. kansasii Sub-Typing by PRA-Hsp65 and rpoB Gene Sequencing

Number of Isolates	BstE II Patterns	Hae III Patterns	Best Matches by PRA	Sequence Typing
24	235, 210	130, 105, 80	M. kansasii type I	M. kansasii type I
8	235, 130, 85	130, 105	M. kansasii type II	M. kansasii type II
1	235, 120, 85	130, 115, 75, 60	M. kansasii type IV	M. kansasii type IV

Figure 2 A dendrogram (the maximum likelihood tree generated using the Tamura 3-parameter model in MEGA6 software) illustrating the genetic distances between different M. kansasii types (I–VI), based on the sequencing results of the partial ropB gene.