Table 1.
Separation of 100 M. abscessus subsp. abscessus clinical strains into genotype level by sequence analyses based on the partial hsp65 (603 bp) and rpoB (711 bp) gene sequences.
Fig 1.
Phylogenetic trees based on the partial hsp65 and rpoB gene sequences of two Rec-mas-H strains.
Phylogenetic trees of 2 Rec-mas-H strains based on (A) the partial sequence of the hsp65 gene (603 bp) and (B) the partial sequence of the rpoB gene (711 bp). The trees were constructed using the neighbor-joining method in the MEGA 4.0 program. The bootstrap values were calculated from 1,000 replications; values <50% are not shown. The black-centered circles indicate that the corresponding clusters were supported with maximum parsimony-based trees. The bar indicates the number of base substitutions per site. The black-centered triangles indicate that the corresponding sequences were sequenced and obtained in this study.
Fig 2.
Neighbor-joining phylogenetic trees based on the 7 MLST genes of two Rec-mas-H strains.
Phylogenetic trees of two Rec-mas-H strains constructed based on the partial sequencing of six MLST genes. (A) argH, (B) cya, (C) glpK, (D) gnd, (E) murC, (F) pta and (G) purH gene sequence based trees were constructed using the neighbor-joining method in the MEGA 4.0 program. The bootstrap values were calculated from 1,000 replications; values <50% are not shown. The black-centered circles indicate that the corresponding clusters were supported by maximum parsimony-based trees. The bar indicates the number of base substitutions per site.
Table 2.
Comparison of two Rec-mas-H strains with reference strains of M. abscessus group in sequence similarities of 7 MLST, hsp65 (603 bp) and rpoB (711 bp) gene sequences.
Fig 3.
Neighbor-joining phylogenetic trees based on the concatenated sequences of two Rec-mas-H strains.
Phylogenetic trees based on (A) concatenation of 7 MLST gene sequences, (B) concatenation of 7 MLST gene sequences and the hsp65 gene sequence, and (C) concatenation of 7 MLST gene sequences and the hsp65 and rpoB gene sequences. The trees for all studied strains were generated using the neighbor-joining method. The bootstrap support values (%) from 1,000 replications are indicated for each node; values <50% are not shown. The black-centered circles indicate that the corresponding clusters were supported by maximum parsimony-based trees. The bar indicates the number of base substitutions per site.
Fig 4.
Identification of two Rec-mas-H strains at the subspecies level by PCR targeting the erm(41) gene.
M, 100-bp DNA ladder; Lane 1, M. abscessus subsp. abscessus CIP 104536T; Lane 2, M. abscessus subsp. massiliense CIP 108297T; Lane 3, M. abscessus subsp. bolletii CIP 108541T; Lane 4, Asan 55814; Lane 5, Asan 55262; N, negative control.
Fig 5.
Differentiation of two Rec-mas-H strains by PCR-restriction enzyme and polymorphism analysis (PRA) of hsp65.
Amplified hsp65 gene amplicons were digested with (A) BstEII and (B) HaeIII restriction enzymes. M, 100-bp DNA ladder; Lane 1, M. abscessus subsp. abscessus CIP 104536T; Lane 2, M. abscessus subsp. massiliense CIP 108297T; Lane 3, Asan 55814; Lane 4, Asan 55262.