Figure 1.
Sequence polymorphisms between the three hsp65 sequevars of M. massiliense.
Type I strains have the same sequence as the M. massiliense type strain; however, Type II-1 and Type II-2 strains differed from the M. massiliense type strain by 2-bp (T444A and C714A) and 1-bp (T444A), respectively. The nucleotide numbers correspond to those from the complete sequence of the heat shock protein 65 kDa (hsp65) gene of M. abscessuss ATCC 19977 (GenBank no. CU458896).
Figure 2.
Phylogenetic trees based on the hsp65 gene (603 bp) and rpoB gene (711 bp) sequences.
Phylogenetic trees based on the (A) hsp65 gene (603 bp) and (B) rpoB gene (711 bp) sequences from M. massiliense clinical isolates, M. massiliense CIP 108297T, M. bolletii CIP 108541T, M. chelonae ATCC 19237, and M. abscessus ATCC 19977. These trees were constructed using the neighbor-joining method. The bootstrap values were calculated from 1,000 replications. Bootstrap values of <50% are not shown. The bars indicate numbers of substitutions per nucleotide position.
Table 1.
Separation of 109 M. abscessus related Korean strains used in this study into species or genotype level by sequence analysis based on the partial hsp65 gene sequence (603 bp).
Table 2.
The frequency of the two hsp65 genotypes (Type I and Type II) determined by hsp65 sequence analysis and Hinf I PRA methods, and the two colony morphotypes (rough and smooth) among 65 M. massiliense clinical strains.
Table 3.
The frequency of the four rpoB genotypes (R-I to R-IV) determined by rpoB sequence analysis (711-bp) among 65 M. massiliense clinical strains, polymorphisms of rpoB genotypes and relationships between rpoB and hsp65 genotypes.
Figure 3.
Colony morphology and the growth patterns in 7H9 broth medium.
Colony morphology (left panel) and the growth patterns in 7H9 broth medium (right panel) of (A) M. massiliense CIP 108297T, (B) M. massiliense Type I strain, and (C) M. massiliense Type II strain.
Figure 4.
Identification of M. massiliense Type I and Type II strains by hsp65 PRA method.
(A) Hinf I PRA algorithm for differentiating of M. massiliense Type I and Type II strains. (B) Agarose gel electrophoresis after Hinf I PRA. Lanes: M, 100-bp ladder; 1, M. abscessus (uncut); 2, M. abscessus (Hinf I cut); 3, M. bolletii (Hinf I cut); 4, M. chelonae (Hinf I cut); 5, M. fortuitum (Hinf I cut); 6, M. massiliense (Hinf I cut); 7, 50375 (Type I, uncut); 8, 50375 (Type I, Hinf I cut); 9, 51302 (Type I, Hinf I cut); 10, 51843 (Type I, Hinf I cut); 11, 52352 (Type I, Hinf I cut); 12, 51048 (Type II, uncut); 13, 51048 (Type II, Hinf I cut); 14, 52008 (Type II, Hinf I cut); 15, 50594 (Type II, Hinf I cut); 16, 52012 (Type II, Hinf I cut).
Figure 5.
MALDI-TOF mass spectrometry analysis.
MALDI-TOF mass spectrometry analysis of extracted lipids from (A) M. massiliense CIP 108297T, (B) 50375 (Type I), (C) 51843 (Type I), (D) 50594 (Type II), and (E) 51048 (Type II). DG, diglycosylated GPLs; TG, triglycosylated GPLs.