Detection of Campylobacter jejuni in food with TaqMan-MGB probe based on real-time PCR assay

Objective:To establish a real-time PCR assay based on TaqMan-MGB probe for detection of Campylobacter jejuni isolated from foods to provide a rapid and accurate tool for monitoring food contamination.Methods:The primers and probe from conserved genomic sequence were designed based on TaqMan-MGB principle.The 5′ end of probe was labeled with FAM fluoresceins,and then the 3′ end of probe was labeled with MGB.The specificity,sensitivity and reproducibility of real-time PCR based on TaqMan-MGB probe were estimated.Finally,this assay was applied to detected specimens collected from market.Results:Campylobacter jejuni was idengtified by using real-time PCR combined with TaqMan-MGB probe accurately and quickly.Then the detection limit of this method for pure culture of Campylobacter jejuni was 10 cfu/ml.When this assay was applied directly to identify 50 clinical specimens,the results showed that 6 were positive to Campylobacter jejuni.The above results were the same to the results obtained from the conventional assays.Conclusion:It is demonstrated that real-time PCR combined with TaqMan-MGB probe reported here is a reliable,accurate and feasible assay and more sensitive than the conventional method.It can be applied to investigation of food poisoning and clinical inspection.