Aims: To develop a rapid multiplex PCR method for simultaneous detection of
five major foodborne pathogens (Staphylococcus aureus, Listeria monocytogenes,
Escherichia coli O157:H7, Salmonella Enteritidis and Shigella flexneri, respectively).
Methods and Results: Amplification by PCR was optimized to obtain high efficiency.
Sensitivity and specificity assays were investigated by testing different
strains. With a multipathogen enrichment, multiplex PCR assay was able to
simultaneously detect all of the five organisms in artificially contaminated pork
samples. The developed method was further applied to retail meat samples, of
which 80% were found to be positive for one or more of these five organisms.
All the samples were confirmed by traditional culture methods for each individual
species.
Conclusions: This study reported a rapid multiplex PCR assay using five primers
sets for detection of multiple pathogens. Higher consistency was obtained
between the results of multiplex PCR and traditional culture methods.
Significance and Impact of the Study: This work has developed a reliable, useful
and cost-effective multiplex PCR method. The assay performed equally as
well as the traditional cultural method and facilitated the sensitive detection
both in artificially contaminated and naturally contaminated samples.