This application note describes the 5’-Nuclease PCR Assays for Foodborne Pathogen Detection using the Agilent-Stratagene Mx3000P Q-PCR System. This method is rapid, highly sensitive and specific.
The text describes a study detailing the use of 5'-nuclease PCR (Q-PCR) as a rapid alternative to conventional culture techniques for detecting pathogenic microbial contaminants in food. The assays specifically target genes from Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, and Campylobacter jejuni.
The methodology involves using primers and probes designed to detect specific genomic DNA sequences. Assays were conducted on the Agilent-Stratagene Mx3000P thermal cycler system with Brilliant Q-PCR Master Mix and analyzed using MxPro software. The study reports detection limits between 1-10 fg of genomic DNA per reaction and a linear quantification range over 6-7 log units.
The assays are designed to detect Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, and Campylobacter jejuni.
Results can be obtained with these assays in 2.5 h following enrichment.
The Campylobacter assay has a detection limit of 1 fg/rxn.