Development of a Multiplex-PCR Serotyping Assay for Characterizing Legionella pneumophila Serogroups Based on the Diversity of Lipopolysaccharide Biosynthetic Loci

Legionella pneumophila, which is the main cause of Legionnaires' disease, comprises at least 15 serogroups (SGs). We show here the diversity of lipopolysaccharide biosynthetic loci among serogroups and describe the development of a PCR serotyping assay for 15 SGs based on the sequences of LPS b...

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Published inJournal of clinical microbiology Vol. 59; no. 11; p. e0015721
Main Authors Nakaue, Ryota, Qin, Tian, Morita, Masatomo, Ren, Hongyu, Chang, Bin, Murai, Miyo, Amemura-Maekawa, Junko, Ohnishi, Makoto
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 19.10.2021
Subjects
Bacteriology
Clinical Microbiology
Humans
Legionella
Legionella pneumophila - genetics
Legionnaires' Disease - diagnosis
Lipopolysaccharides
Multiplex Polymerase Chain Reaction
Serogroup
Serotyping
serogroup
Legionella pneumophila
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Summary:Legionella pneumophila, which is the main cause of Legionnaires' disease, comprises at least 15 serogroups (SGs). We show here the diversity of lipopolysaccharide biosynthetic loci among serogroups and describe the development of a PCR serotyping assay for 15 SGs based on the sequences of LPS biosynthetic loci. Using this multiplex-PCR (M-PCR) system, serogroups were detected using primers that specifically amplify the sequences of SG1, SG2, SG5, SG7, SG8, SG9, SG11, SG13, SG3/15, and SG6/12. When PCR products of the expected sizes were not detected, we used primers that identified SG4/10/14. The PCR serotyping system specifically amplified the sequences corresponding to SGs of 238 L. pneumophila strains. This method will be very useful for conducting epidemiological studies and investigating outbreak of Legionnaires' disease.
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Ryota Nakaue and Tian Qin contributed equally to this work. Author order was determined alphabetically.
Present address: Ryota Nakaue, Department of Clinical Laboratory, The University of Tokyo Hospital, Tokyo, Japan.
Citation Nakaue R, Qin T, Morita M, Ren H, Chang B, Murai M, Amemura-Maekawa J, Ohnishi M. 2021. Development of a multiplex-PCR serotyping assay for characterizing Legionella pneumophila serogroups based on the diversity of lipopolysaccharide biosynthetic loci. J Clin Microbiol 59:e00157-21. https://doi.org/10.1128/JCM.00157-21.
ISSN:0095-1137
1098-660X
DOI:10.1128/jcm.00157-21