Detection of Campylobacter jejuni by Fluorescent in Situ Hybridization

Hazard Analysis Critical Control Point (HACCP) systems require verification procedures that include confirmation of the presence of pathogens in finished product and processing environments. C. jejuni is a microaerophilic organism that requires specialized culture conditions for growth. This involves the use of special gaseous atmospheres and culture supplements. Thus, only a few specialized laboratories can currently analyze food and environmental samples for C. jejuni and testing tends to be very expensive. This significantly impedes monitoring and surveillance of C. jejuni in poultry production.
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This project will use the molecular biology approach known as fluorescent in situ hybridization to develop a rapid, culture independent procedure for the detection of C. jejuni in poultry and in environmental samples from poultry processing environments. This approach will overcome the difficulties associated with culturing while still being specific for viable C. jejuni. The work will be completed in three milestones: first, the specificity of an existing fluorescent in situ hybridization (FISH) protocol will be confirmed using a wide range of C. jejuni isolates and non-Campylobacter isolates. Second, a buoyant centrifugation based procedure will be adopted which will allow to concentrate C. jejuni from large sample volumes and to separate microbial cells from particles that can interfere with FISH detection. Third, the newly established FISH protocol will be tested and validated with spiked and naturally contaminated poultry products and environmental samples from processing environments.