The Campylobacter jejuni real-time PCR rapid detection kit was developed， and validated according to the AOAC microbiology method validation guidelines. The specificity of the kit was 100% for the 115 target strains and 118 non-target strains， and the sensitivity reached 6.4 CFU/mL. The Ct values for repeated freezing and thawing for 30 times didn't show significant differences. Robutstness results showed that the enrichment broth volume， heating lysis time and the DNA volume in PCR reaction didn't have any influence on the PCR result for the 11 Campylobacter jejuni and 5 non-target Campylobacter strains， except for one Campylobacter jejuni strain， for which high volume of enrichment broth can inhibit PCR reaction. The combined lot-to-lot and stability test showed the three lots （newly produced， 4 months after production， 1 year after production） gave consistent results for the 16 strains， and the Ct values for the 11 Campylobacter jejuni were similar. Consistent results were shown both for the kit method and conventional method in the innoculation of Campylobacter jejuni in chicken， vegetable and milk samples， as well as in practical chicken sample detection， and there was no “false negative” using the kit method. Our kit is open， can be used on various real-time PCR platforms， contained the internal amplification control which can avoid the “false negative result”， and can be applied in the rapid detection of Campylobacter jejuni from food.