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Abstract
Twenty two strains of Legionella species isolated from Jejudo, Korea were identified by comparing the rpoB (300 bp), dotA (360 bp), and mip (396 bp) gene sequence analysis. Furthermore, their genotypes were determined by sequence analysis of rpoB/dotA subgroup typing, pulse-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) patterns. Of the 22 isolates, 21 strains were identified as L. pneumophila and 1 strain was close to L. erythra (>95% similarity of rpoB and mip). Most of the L. pneumophila strains (90%) belonged to P-I of rpoB/dotA subgroup typing, one strain of each P-III and P-IV. L. pneumophila isolates were further grouped into 4 and 6 different PFGE (P1 to P4) and RAPD (R1 to R6) patterns, respectively. On the basis of these genotypes, which may be useful for future epidemiological studies, existence of diverse L. pneumophilla population in Jejudo, Korea were observed.
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Figure 1.
Genetic relationships of Legionella Jeju isolates (•) among 41 legionella species based on the rpoB and mip sequences. In addition to the sequence similarity, most of the Jeju isolates were clearly identified as L. pneumophila, except one strain (Jeju 8-5). Strains possessing sequences identical to those in the figures are not presented. Bootstrap values higher than 50% are shown.
Figure 2.
Neighbour-joining trees inferred from (A) the rpoB and (B) the dotA sequences of L. pneumophila Jeju isolates and reference strains (K and JLP) (26). Subgroups (P-I to PVIII and F-I and F-II) are indicated on the right; branch lengths are proportional to the number of nucleotide changes. Bootstrap values higher than 50% are indicated at the corresponding branches. SG, serogroup numbers (SG 1 to 15).
Figure 3.
PFGE patterns of SfiI-cleaved genomic DNAs from L. pneumophila Jeju isolates (A) and their schematic four band patterns (PG1 to PG4) (B). Lambda ladder PFGE marker (M) were used as molecular size standards.
Figure 4.
RAPD typing of L. pneumophila Jeju isolates. Six different band patterns (Type R1 to R6) were observed (B). ATCC 33152, L. pneumophila serogroup 1; M, 100-bp ladder marker; N, negative control.
Table 1.
Primers to amplify partial fragments of target genes of Legionella isolates
Table 2.
Distribution of L. pneumophila Jeju isolates based on the rpoB and dotA sequences, and RAPD and PFGE patterns
| Isolates | Subgroup | PR type | Serogroup | ||
|---|---|---|---|---|---|
| rpoB | dotA | rpoBa | dotAa | ||
| Jeju 1-1 | P-I | P-I | P3 R1 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 1-2 | P-I | P-I | P3 R5 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 1-3 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 2 | P-I | P-I | P2 R1 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 3-1 | P-I | P-I | P3 R3 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 3-2 | P-I | P-I | P3 R3 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 4-1 | P-I | P-I | P2 R1 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 4-2 | P-I | P-I | P2 R1 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 4-3 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 5-3 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 6-1 | P-I | P-I | P2 R1 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 6-2 | P-I | P-I | P2 R4 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 6-3 | P-III | P-III | P4 R6 | SG 2, SG 6, SG 12 | SG 12 |
| Jeju 6-5 | P-I | P-I | P2 R2 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 6-6 | P-I | P-I | P2 R2 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 6-7 | P-I | P-I | P1 R4 | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 7-1 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 8-1 | P-IV | P-IV | ND | SG 7, SG 14 (99.3) | SG 14 (97.5) |
| Jeju 8-2 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 8-3 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
| Jeju 8-4 | P-I | P-I | ND | SG 1-4 | SG 1-1, SG 1-4, SG 9 |
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