ABSTRACT
OBJECTIVE@#To investigate the molecular characteristics and intracellular growth ability of Legionella pneumophila (L. pneumophila) strains from 1989 to 2016 in Sichuan Province, China.@*METHODS@#Seventy-nine isolates of L. pneumophila were collected from environmental and clinical sources, including cooling towers, hot springs, bath water, fountains, and patients, and identified with 16S rRNA gene analysis and serum agglutination assay. The isolates were then typed by Sequence-Based Typing (SBT), and Genotyping of forty-two LP1 strains were analyzed by means of multiple-locus VNTR analysis with 8 loci (MLVA-8). All strains were further analyzed for two virulence genes: Legionella vir homologue (lvh) and repeats in structural toxin (rtxA). The intracellular growth ability of 33 selected isolates was determined by examining their interaction with J774 cells.@*RESULTS@#All isolates were identified to L. pneumophila including 11 serogroups, among which the main serogroup were LP1, accounting for 54.43%. Thirty-three different sequence types (STs) from five main clonal groups and five singletons were identified, along with 8 different MLVA patterns. Both the lvh and rtxA loci were found in all 79 strains. Thirty isolates showed high intracellular growth ability in J774 cells.@*CONCLUSION@#L. pneumophila is a potential threat to public health, and effective control and prevention strategies are urgently needed.
Subject(s)
Humans , Bacterial Proteins , Genetics , Bacterial Toxins , Genetics , China , Genotyping Techniques , Legionella pneumophila , Genetics , RNA, Ribosomal, 16S , Genetics , Water MicrobiologyABSTRACT
In this study,the L.dumoffii TEX-KL (ATCC 33343),L.dumoffii NY23 (ATCC 33279) and L.pneumophila philadelphila-1 (ATCC 33155) strains were used to explore the adhesion,invasion and intracellular growth ability in the epithelial cells.Approximately 1× 108 bacteria were pelleted,resuspended,and diluted (1 ∶ 10) in RPMI 1640 tissue culture medium.The bacteria were then added to A549 cells (1 × 105 per well) in 24-well plates to give a multiplicity of infection (MOI) of about 100.The Gimenez staining and colony counting methods were used for the determination of the strain adhesion,invasion and intracellular growth ability.It was found that in vitro growth ability of L.pneumophila philadelphila-1,L.dumoffii TEX-KL and L.dumoffii NY23 strains had no significant difference.For in vivo assay,there was also no significant difference in adhesion ability of these strains.However,the CFU counts of L.dumoffii TEX-KL strain invaded into A549 cells was 1 000 times higher than that of the other two strains.Compared with L.pneumophila philadelphila-1 and L.dumoffii NY23 strains,L.dumoffii TEX-KL strain has higher invasion ability and,therefore,higher intracellular growth ability.
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.