Figure 1Phylogenetic trees based on the hsp65 and rpoB sequence types in the clinical isolates of Mycobacterium intracellulare. Phylogenetic analysis of the (A) hsp65 sequence type (B) rpoB sequence type.
Figure 2Comparison of the dendrograms from the cluster analysis of Mycobacterium intracellulare using categorical coefficients.
Figure 3Dendrogram of the cluster analysis based on DNA restriction banding patterns of the Mycobacterium intracellulare clinical isolates using SpeI.
Figure 4Cluster analysis of Mycobacterium intracellulare based on variable-number tandem-repeat analysis profiles. *M0016: Reference strain ATCC 13950.
Figure 5Cluster analysis of Mycobacterium intracellulare based on mycobacteria interspersed repetitive-unit-variable-number tandem-repeat analysis profiles. *M0016: Reference strain ATCC 13950.
Table 1Primers used for multilocus sequence typing in this study.
Gene |
Primer |
Sequences (5′–3′) |
Amplicon size (bp) |
No. of alleles |
argH
|
argH-F |
AGTCGTGCAGGGTGACGTAG |
495 |
11 |
argH-R |
CCACCCTGTTCCGGATGT |
cya
|
cya-F |
CCGTGCGGATCCTTCAGT |
660 |
7 |
cya-R |
GTGCTGTTGCTGGGCATC |
glpK
|
glpK-F |
ATCGGATCGCCAGTAGGG |
670 |
6 |
glpK-R |
GACACCTGGGTGTTGTGGAA |
gnd
|
gnd-F |
TGGCGGTCACCACGACAC |
555 |
4 |
gnd-R |
GATCACGGCGCAGGTCTT |
murC
|
murC-F |
GTTGCTCGGGGTCCAGGT |
607 |
4 |
murC-R |
GTCGTCGCTGGACCTGTTG |
pta
|
pta-F |
GGTTCGATGCCGAATTGTG |
665 |
8 |
pta-R |
CACGTCGACATCACCGATCT |
Table 2Primers used for the mycobacteria interspersed repetitive-unit-variable-number tandem-repeat analysis in this study.
Set |
Alias |
PCR primer pairs (5′–3′, labeling indicated) |
Primer concentration |
Set 1 |
MIN18 |
GCCGAACCATTTGGCGAAC(VIC) |
4μM |
|
|
GGATTCGGCCGCGCAATTC |
|
|
MIN19 |
CATGGTTCGCCCTCTACAC(PET) |
5μM |
|
|
TAGGGGCAGGTCATCGAAG |
|
|
MIN20 |
GCTGAGCTACAGCCTCGAC(NED) |
7.5μM |
|
|
CGACGCCGATGACGTAAAC |
|
Set 2 |
MIN22 |
TCAGGAATGGGTCCGGTTC(FAM) |
5μM |
|
|
AGCTCGTGACGACGGAAAC |
|
|
MIN31 |
CGACCGCATCCAGAAACAG(VIC) |
5μM |
|
|
GCTCTATGACGACCTCAAG |
|
|
MIN33 |
GTGCAGTTCAACCACGAAC(PET) |
7.5μM |
|
|
GGCGTTGAACACGTTGGTG |
|
Table 3Type distribution of antibacterial susceptibility analysis in the clinical isolates of Mycobacterium intracellulare.
Antibiotic susceptibility type |
Antibacterial drug
|
AS Type No. (%) |
CLA |
MXF |
RIF |
SXT |
AMI |
LZD |
CIP |
DOX |
AST1 |
S |
R |
R |
R |
S |
R |
R |
R |
28 (63.6) |
AST2 |
S |
R |
R |
S |
S |
R |
R |
R |
7 (15.9) |
AST3 |
S |
R |
R |
R |
R |
R |
R |
R |
4 (9.1) |
AST4 |
S |
S |
S |
R |
S |
S |
S |
S |
2 (4.5) |
AST5 |
S |
R |
R |
S |
R |
R |
R |
R |
1 (2.3) |
AST6 |
S |
S |
S |
S |
S |
S |
S |
S |
1 (2.3) |
AST7 |
S |
S |
S |
R |
S |
R |
S |
S |
1 (2.3) |
Table 4VNTR and MIRU-VNTR allele distribution and allelic diversity of 44 Mycobacterium intracellulare clinical isolates.
Methods |
Locus |
No. of alleles in 44 Mycobacterium intracellulare clinical isolates
|
Allelic diversity (h) |
0 |
1 |
2 |
3 |
4 |
5 |
6 |
VNTR |
VNTR1 |
— |
— |
42 |
2 |
— |
— |
— |
0.089 |
VNTR2 |
— |
30 |
8 |
4 |
2 |
— |
— |
0.503 |
VNTR3 |
3 |
40 |
1 |
— |
— |
— |
— |
0.172 |
VNTR4 |
— |
26 |
5 |
5 |
8 |
— |
— |
0.606 |
VNTR5 |
10 |
— |
34 |
— |
— |
— |
— |
0.359 |
VNTR6 |
1 |
— |
19 |
24 |
— |
— |
— |
0.527 |
VNTR7 |
1 |
16 |
14 |
12 |
1 |
— |
— |
0.707 |
VNTR8 |
22 |
— |
22 |
— |
— |
— |
— |
0.512 |
VNTR9 |
4 |
17 |
21 |
2 |
— |
— |
— |
0.627 |
VNTR10 |
— |
31 |
2 |
— |
10 |
— |
— |
0.458 |
VNTR12 |
— |
1 |
31 |
12 |
— |
— |
— |
0.439 |
VNTR13 |
— |
— |
27 |
17 |
— |
— |
— |
0.485 |
VNTR14 |
— |
1 |
41 |
2 |
— |
— |
— |
0.132 |
VNTR15 |
— |
29 |
10 |
5 |
— |
— |
— |
0.513 |
VNTR16 |
— |
— |
44 |
— |
— |
— |
— |
0 |
MIRU-VNTR |
MIN18 |
— |
— |
— |
— |
9 |
31 |
4 |
0.464 |
MIN20 |
— |
— |
11 |
31 |
2 |
— |
— |
0.449 |
MIN19 |
— |
— |
16 |
14 |
14 |
— |
— |
0.681 |
MIN22 |
— |
— |
32 |
3 |
9 |
— |
— |
0.434 |
MIN31 |
— |
— |
35 |
9 |
— |
— |
— |
0.333 |
MIN33 |
— |
— |
— |
26 |
16 |
2 |
— |
0.529 |