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

J Agric Food Chem. 2022 Aug 3;70(30):9557-9566. doi: 10.1021/acs.jafc.2c02581. Epub 2022 Jul 20.

Abstract

Campylobacter jejuni 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 C. jejuni in food samples. The two assays were found to be highly specific to C. jejuni 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 C. jejuni 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 C. jejuni contamination in food samples, particularly for work done in the field or poorly equipped labs.

Keywords: CRIPSR/Cas12a; Campylobacter jejuni; exo-probe; food samples; recombinase-aided amplification.

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Campylobacter jejuni* / genetics
  • Cattle
  • Chickens
  • Food Analysis*
  • Nucleic Acid Amplification Techniques / methods
  • Recombinases / genetics

Substances

  • Recombinases