Abstract
BackgroundCholera is an acute diarrheal disease caused by Vibrio cholerae. Outbreaks are caused by a genetically homogenous group of strains from serogroup O1 or O139 that are able to produce the cholera toxin. Rapid detection and identification of these epidemic strains is essential for an effective response to cholera outbreaks.ResultsThe use of ferulic acid as a matrix in a new MALDI-TOF MS assay increased the measurable mass range of existing MALDI-TOF MS protocols for bacterial identification. The assay enabled rapid discrimination between epidemic V. cholerae O1/O139 strains and other less pathogenic V. cholerae strains. OmpU, an outer membrane protein whose amino acid sequence is highly conserved among epidemic strains of V. cholerae, appeared as a discriminatory marker in the novel MALDI-TOF MS assay.ConclusionsThe extended mass range of MALDI-TOF MS measurements obtained by using ferulic acid improved the screening for biomarkers in complex protein mixtures. Differences in the mass of abundant homologous proteins due to variation in amino acid sequences can rapidly be examined in multiple samples. Here, a rapid MALDI-TOF MS assay was developed that could discriminate between epidemic O1/O139 strains and other less pathogenic V. cholerae strains based on differences in mass of the OmpU protein. It appeared that the amino acid sequence of OmpU from epidemic V. cholerae O1/O139 strains is unique and highly conserved.
Highlights
Cholera is an acute diarrheal disease caused by Vibrio cholerae
Forty-eight isolates acquired from different strain collections (Table 1) and previously identified as V. cholerae were analyzed using MALDI-TOF MS and Biotyper 2.0 software (Bruker Daltonics)
We demonstrate that the outer membrane protein OmpU from V. cholerae can be used as a biomarker of epidemic strains of V. cholerae in a new adapted MALDI-TOF MS assay
Summary
Cholera is an acute diarrheal disease caused by Vibrio cholerae. Outbreaks are caused by a genetically homogenous group of strains from serogroup O1 or O139 that are able to produce the cholera toxin. Rapid detection and identification of these epidemic strains is essential for an effective response to cholera outbreaks. Cholera is an acute diarrheal disease caused by Vibrio cholerae that can be lethal within hours if left untreated. The severity, duration, and frequency of cholera epidemics appear to be increasing [2], indicating that cholera is a severe public health problem. V. cholerae is considered a category B bioterrorism agent by the CDC [3]. Within the V. cholerae species, over 200 serogroups have been identified but only serogroup O1 and O139 strains Infection usually occurs by consumption of contaminated water, the natural habitat of V. cholerae, or contaminated food.
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