A Robust and Poisson Validated Quantitative 5′ Nuclease TaqMan® Real-Time PCR Assay Targeting fimA for the Rapid Detection of Salmonella spp. in Food

A newly designed TaqManA (R) probe and an internal amplification control were implemented in a conventional PCR system targeting the major fimbrial subunit encoding gene fimA. This assay has an inclusivity and exclusivity of 100 % (n = 126). The limit of detection (LOD) and the absolute quantification limit, both determined by advanced Poisson analyses, were three bacterial cell equivalents per quantitative real-time PCR reaction. The fimA assay achieved 100 % accuracy and performed very robustly, producing reproducible, reliable data for quantification of Salmonella spp., even at the LOD.