Development of Recombinase-Aided Amplification (RAA)-Exo-Probe and RAA-CRISPR/Cas12a Assays for Rapid Detection of <i>Campylobacter jejuni</i> in Food Samples

Abstract

<i>Campylobacter jejuni</i> is the major cause of campylobacteriosis, one of the most common foodborne illnesses worldwide. Here, we report the development of RAA-exo-probe and RAA-CRIPSR/Cas12a assays for the detection of <i>C. jejuni</i> in food samples. The two assays were found to be highly specific to <i>C. jejuni</i> and highly sensitive, as they were one log more sensitive compared to the traditional culture method, with detection thresholds of 9 and 5 copies per reaction, respectively. These assays successfully detected <i>C. jejuni</i> in spiked chicken samples and natural meat samples (chicken, beef, mutton, etc.) and were overall less dependent on expensive equipment, only requiring a fluorescent reader. Their ease of use compared to other nucleic acid amplification-based methods indicates that these assays could be adapted for the rapid, routine surveillance of <i>C. jejuni</i> contamination in food samples, particularly for work done in the field or poorly equipped labs.