Bacteriophage tailspike protein based assay to monitor phase variable glucosylations in Salmonella O-antigensReportar como inadecuado

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BMC Microbiology

, 16:207

Applied microbiology


BackgroundNon-typhoid Salmonella Typhimurium S. Typhimurium accounts for a high number of registered salmonellosis cases, and O-serotyping is one important tool for monitoring epidemiology and spread of the disease. Moreover, variations in glucosylated O-antigens are related to immunogenicity and spread in the host. However, classical autoagglutination tests combined with the analysis of specific genetic markers cannot always reliably register phase variable glucose modifications expressed on Salmonella O-antigens and additional tools to monitor O-antigen glucosylation phenotypes of S. Typhimurium would be desirable.

ResultsWe developed a test for the phase variable O-antigen glucosylation state of S. Typhimurium using the tailspike proteins TSP of Salmonella phages 9NA and P22. We used this ELISA like tailspike adsorption ELITA assay to analyze a library of 44 Salmonella strains. ELITA was successful in discriminating strains that carried glucose 1-6 linked to the galactose of O-polysaccharide backbone serotype O1 from non-glucosylated strains. This was shown by O-antigen compositional analyses of the respective strains with mass spectrometry and capillary electrophoresis. The ELITA test worked rapidly in a microtiter plate format and was highly O-antigen specific. Moreover, TSP as probes could also detect glucosylated strains in flow cytometry and distinguish multiphasic cultures differing in their glucosylation state.

ConclusionsTailspike proteins contain large binding sites with precisely defined specificities and are therefore promising tools to be included in serotyping procedures as rapid serotyping agents in addition to antibodies. In this study, 9NA and P22TSP as probes could specifically distinguish glucosylation phenotypes of Salmonella on microtiter plate assays and in flow cytometry. This opens the possibility for flow sorting of cell populations for subsequent genetic analyses or for monitoring phase variations during large scale O-antigen preparations necessary for vaccine production.

KeywordsSalmonella Typhimurium O-antigen Tailspike protein Bacteriophage Phase variation O-serotyping Flow cytometry AbbreviationsCE-LIFCapillary electrophoresis with laser-induced fluorescence detection

ELITAELISA like tailspike adsorption

HPAEC-PADHigh performance anion exchange with pulsed amperometric detection


MALDI-MSMatrix-assisted laser-desorption ionization mass spectrometry

TSPTailspike protein.

Electronic supplementary materialThe online version of this article doi:10.1186-s12866-016-0826-0 contains supplementary material, which is available to authorized users.

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Autor: Andreas Schmidt - Wolfgang Rabsch - Nina K. Broeker - Stefanie Barbirz


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