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Development and evaluation of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species
EP31544
Poster Title: Development and evaluation of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species
Submitted on 27 Apr 2020
Author(s): Hongsheng Huang and Beverley Phipps-Todd
Affiliations: Canadian Food Inspection Agency, Ottawa Laboratory – Fallowfield ,Ottawa, Ontario, Canada K2H 8P9
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Poster Information
Abstract: Campylobacter species, particularly thermotolerant Campylobacter spp., such as C. jejuni, are major human foodborne pathogens. Culture methods have been routinely used for the detection of this organism in various types of samples. An alternative, simple and rapid confirmation test(s) without further tedious biochemical tests would be useful. Meanwhile, Campylobacter-like colonies can be difficult to identify on agar plates overgrown with competitive bacteria, which can lead to false-negative results. This study developed a simple colony blot immunoassay using a new monoclonal antibody (Mab) produced in the present study for rapid screening, confirmation and quantification of campylobacters on culture agar plates. The procedure developed in this study was able to specifically detect thermotolerant Campylobacter spp., but not other non-thermotolerant Campylobacter and non-Campylobacter reference strains tested. This assay could detect 100000 Campylobacter cells in a single dot. For 39 membranes transferred from agars cultured with natural chicken meat rinse samples, the CBI showed 100% correlation (16 positive and 23 negative for C. jejuni ) with cultural tests. The assay developed in this study can be used as quantitative and qualitative diagnostic test to identify and confirm the presence of thermotolerant Campylobacter (including C. jejuni, C. coli, C. lari and C. upsaliensis) on agar plates. Summary: Cultural procedures to detect foodborne thermotolerant campylobacters are tedious. It is often difficult to differentiate Campylobacter colonies from non-Campylobacter colonies growing on the same agar plate. This poster showed a simple colony blot immunoassay as an alternative procedure to facilitate the rapid screening, specific identification and quantification of Campylobacter colonies on agar plates with or without the overgrowth of competitive bacteria to avoid false-negative results.References: Huang H, Phipps-Todd B, McMahon T, Elmgren CL, Lutze-wallace C, Todd ZA, Garcia MM. 2016. Development and evaluation of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species. J Microbial Methods. 130: 76-82.

Huang H, Raymond P, Grenier C, Fahey J. 2020. Development and improvement of a colony blot immunoassay for the detection of thermotolerant Campylobacter species. Methods in Microbiology 47, Immunological Methods in Microbiology, Elsevier. In press. Available online (https://www.elsevier.com/books/immunological-methods-in-microbiology/gurtler/978-0-12-820811-3). Official publication in May 2020.
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