Development of a Detection Method for Food-borne Campylobacter jejuni Based on Fluorescence Probe-based Recombinase Polymerase Amplification
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Abstract:
In order to achieve rapid and convenient detection of Campylobacter jejuni, a rapid detection method of Campylobacter jejuni based on fluorescence probe-based recombinase polymerase amplification (exo-RPA) was developed. The specificity of the method was determined by exo-PA detection of Campylobacter jejuni and control strains. The gradient-diluted Campylobacter jejuni was used as a template to analyze the sensitivity of the exo-RPA method. The application effect of the exo-RPA method was analyzed through detecting the simulated contaminated samples.The exo-RPA and fluorescence PCR were used to test real food samples respectively, in order to analyze their detection effects. The Campylobacter jejuni exo-RPA method can specifically detect Campylobacter jejuni with a detection sensitivity of 6.0×102 CFU/mL. In the simulated contamination tests, the enrichment broth medium containing 2.5×101 CFU/mL Campylobacter jejuni was tested positive after 24 hours of culture by the exo-RPA assay. The exo-RPA and fluorescence PCR methods led to the same detection results for the 40 samples. The Campylobacter jejuni exo-RPA assay developed in this study has high specificity, sensitivity and anti-interference ability. The method is simple, convenient and rapid, with a good application prospect.
