- Application of Multilocus Sequence Analysis (MLSA) for Accurate Identification of Legionella spp. Isolated from Municipal Fountains in Chengdu, China, Based on 16S rRNA, mip, and rpoB Genes
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Wang Guan , Ying Xu , Da-li Chen , Jia-nan Xu , Yu Tian , Jian-ping Chen
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J. Microbiol. 2012;50(1):127-136. Published online February 27, 2012
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DOI: https://doi.org/10.1007/s12275-012-1243-1
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 Abstract
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Legionellosis (Legionnaires’ disease; LD) is a form of severe
pneumonia caused by species of Legionella bacteria. Because
inhalation of Legionella-contaminated aerosol is considered
the major infection route, routine assessments of potential
infection sources such as hot water systems, air-conditioner
cooling water, and municipal fountains are of great importance.
In this study, we utilized in vitro culture and multilocus
sequence analysis (MLSA) targeting 16S rRNA, mip,
rpoB, and mip-rpoB concatenation to isolate and identify
Legionella spp. from 5 municipal fountains in Chengdu
City, Sichuan Province, China. Our results demonstrated
that 16S rRNA was useful for initial identification, as it
could recognize isolates robustly at the genus level, while
the genes mip, rpoB, and mip-rpoB concatenation could
confidently discriminate Legionella species. Notably, the
three subspecies of L. pneumophila could be distinguished
by the analysis based on rpoB. The serotyping result of
strain CD-1 was consistent with genetic analysis based on
the concatenation of mip and rpoB. Despite regular maintenance
and sanitizing methods, 4 of the 5 municipal fountains
investigated in this study were positive for Legionella
contamination. Thus, regularly scheduled monitoring of
municipal fountains is urgently needed as well as vigilant
disinfection. Although the application of MLSA for inspection
of potential sites of infection in public areas is not
standard procedure, further investigations may prove its
usefulness.
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Citations
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