The Microbiologique SCS Real-time PCR Assay utilizes real-time PCR to amplify and detect Cronobacter and Salmonella genomic sequences and an internal control sequence. The assay includes primers and TaqMan® probes for amplification and detection of five different targets. The real-time PCR amplifies three Cronobacter targets, two detected as FAM signal for Cronobacter spp. and one as HEX signal specific for Cronobacter sakazaki. The PCR also amplifies one Salmonella target as ROX and the internal control target as Cy5. The enriched food or environmental swab sample is lysed to release the DNA from cells using a proprietary lysis buffer. The released nucleic acids are then combined with amplification reagents and transferred to a real-time PCR machine for amplification and fluorescence detection. The target bacterial DNA sequences are unique and are not present in other bacteria. The target-specific TaqMan® probes bind to the target amplicons and become cleaved by Taq polymerase during amplification, generating fluorescent signals upon fluorophore and quencher separation. The real-time PCR machine detects the fluorescent signals which, with each PCR cycle proportionately increases as amplicons accumulate.