Study on multiplex PCR assay for detection of Campylobacter jejuni,Listeria monocytogenes and Escherichia coli O157

Objective To establish a multiplex PCR method and apply multiplex PCR for simultaneous detection of Campylobacter jejuni,Listeria monocytogenes and Escherichia coli O157.Methods According to DNA sequences of the mapA gene of Campylobacter jejuni,the hly gene of Listeria monoytogenes and the rfbE gene of Escherichia coli O157,three pairs of specific primers were screened and designed to compose optimized system and conditions of multiplex PCR.The three expected sizes were 589bp for Campylobacter jejuni,360bp for Listeria monocytogenes and 194bp for Escherichia coli O157.Results The detection sensitivity of multiplex PCR were 2.264ng、 37.92ng and 2.100ng for Campylobacter jejuni,Listeria monocytogenes and Escherichia coli O157,respectively.The multiplex PCR took 6-8 hours to detect a sample.Conclusions The results of the experiments demonstrated that the multiplex PCR assay was rapid,simple,sensitive and specific,Which establish important foundation for simultaneous detection for these three bacteria in polluted sample.