Objective To establish a nested polymerase chain reaction(Nested-PCR) assay for rapid detection of Campylobacter jejuni(C.jejuni) in laboratory animals.Method Two pairs of primers were designed according to the flaA gene of C.jejuni.DNA was extracted from C.jejuni.The specific amplified fragment of flaA gene which encoding flagellin A protein of C.jejuni was cloned into the pGEM-T vector and transformed E.coli DH5.The recombinant plasmid was identified by restriction endonuclease analysis,and confirmed by sequencing.Non-C.jejuni and 121 clinical specimens were submitted to detect C.jejuni by nested-PCR.Results C.jejuni PCR amplification product of the external primer was 1 719 bp,and nested-PCR amplification product of the internal primer was 640 bp.The sensitivity of nested-PCR was 10 CFU.The whole detection could be finished within 8 h.Non-C.jejuni did not show specific amplified fragment.121 clinical samples taken randomly from laboratory animals were submitted for detection by nested-PCR and bacteria separation method,with a positive detection rate of 25.6%(31/121) and an agreement of 100%.Conclusion These data demonstrated that the nested-PCR assay established here is a specific and sensitive method for rapid detection of C.jejuni in laboratory animals.