The Validation of the SIMUL-qPCR Salmonella Assay for AOAC Research Institute Performance Tested MethodsSM Certification (Certificate No. 042001).

Background: The Simultaneous Multiplex (SIMUL) Real-Time PCR (qPCR) Salmonella Assay is a quick, reliable method for detecting Salmonella species in environmental and food samples. The assay multiplexes several targets in one run to identify Salmonella species. For most matrixes, the assay uses buffered peptone water for enrichment purposes. However, the assay can be used in conjunction with the SIMUL-qPCR Top 7 STEC Assay for raw beef trim and raw ground beef when proprietary media (Enterohemorrhagic E. coli Recovery and Enrichment Broth or EREB) has been used. This media is optimized for single-step enrichment and recovery of enterohemorrhagic E. coli and Salmonella in those matrixes.
Objective: This report details the method validation study to validate raw beef trim, raw ground beef, fresh raw ground poultry, ready-to-eat cooked poultry, dry pet food, pasteurized liquid eggs, peanut butter, frankfurter/sausage, poultry carcass rinse, and environmental surface sponges and swabs for stainless steel, plastic, rubber, ceramic tile, and sealed concrete.
Method: Matrix studies, inclusivity/exclusivity, product consistency/stability, and robustness testing were conducted to assess the method's performance.
Results: Inclusivity/exclusivity testing showed the SIMUL-qPCR Salmonella Assay was able to detect Salmonella strains while excluding the non-Salmonella isolates. There were no statistically significant differences found between the candidate and reference methods in the matrix studies. Small variations in critical test parameters (enrichment time, extraction reagent volume, and extracted sample volume) didn't adversely affect the assay's performance, and stability testing indicated consistent results for at least one year.
Conclusions: The data presented demonstrate the SIMUL-qPCR Salmonella Assay is a reliable method for detecting Salmonella.
Highlights: The SIMUL-qPCR Salmonella Assay is validated to detect Salmonella in a variety of matrices in as little as 16 hours in Buffered Peptone Water. The assay can also be used in conjunction with the SIMUL Top 7 Assay to test for both Salmonella and the Top 7 Shiga toxin producing E. coli using a single enrichment in EREB media.
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