Development and improvement of a colony blot immunoassay for the detection of thermotolerant Campylobacter species

Publication date: Available online 17 January 2020Source: Methods in MicrobiologyAuthor(s): Hongsheng Huang, Philippe Raymond, Chris Grenier, Jessica FaheyAbstractCampylobacter species, particularly thermotolerant Campylobacter spp., such as C. jejuni, are major human foodborne pathogens worldwide. Culture methods are routinely used for the detection of this organism. Campylobacter-like colonies can be difficult to identify on agar plates overgrown with competitive bacteria. An alternative, simple and rapid confirmation testing without further tedious biochemical tests would be useful. Previously we developed a colony blot immunoassay (CBI) using a specific monoclonal antibody (Mab M1169) for rapid screening, confirmation and quantification of campylobacters on agar plates. This assay showed 100% correlation with the culture method from 21 food samples experimentally inoculated with the most common thermotolerant campylobacters, and over 100 natural non-intentionally inoculated samples of chicken meat, chicken faeces and vegetables. The Mab 1169 was able to specifically detect the most common thermotolerant Campylobacter spp. Further studies have been conducted to improve the assay procedures with semi-automation and to understand the nature of antigen(s) reacting with this antibody using mass spectrometry analysis. It seems that the antibody recognizes an epitope(s) shared between several proteins including Chaperone GroEL and Flagellin A or an antigen bound to these protein...
Source: Methods in Microbiology - Category: Microbiology Source Type: research