Applied Food Diagnostics has announced three AOAC® Performance Tested Methods^SM approvals for its suite of simultaneous, multiplex real-time polymerase chain reaction (SIMUL-qPCR) assays.

SIMUL-qPCR test kits include multiple targets (multiplex) with an internal amplification control, and each kit utilize the same operational parameters allowing simultaneous detection.

The three SIMUL-qPCR assays granted certification include; one test kit for Salmonella, one test kit for both Listeria spp. and Listeria monocytogenes and a suite of test kits for the “Top 7” Shiga toxin-producing Escherichia coli (STEC), including O157: H7, with stx and eae genes.  

The AOAC-RI PTM evaluations consisted of inclusivity/exclusivity, product consistency, stability, robustness and matrix studies. 

These approvals certify the SIMUL-qPCR assay kits perform equivalent or better than standard reference methods (U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM), U. S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (MLG)) in the detection of the specified target within a variety of food matrices and environmental surfaces. 

The SIMUL-qPCR Salmonella Assay was assigned PTM Certification No. 042001. The validation study demonstrated that the performance of the assay is comparable to the MLG, for raw beef trim, raw ground beef, raw ground poultry, RTE cooked poultry, frankfurters, pasteurized liquid eggs and poultry carcass rinses, and the BAM for dry pet food, peanut butter and environmental surface sponges/swabs. 

The SIMUL-qPCR Top 7 STEC Assay Collection for detection of E. coli O157: H7, O26, O45, O103, O111, O121, O145, Shiga toxin and intimin genes were assigned Certification No. 022001. This validation study demonstrated that the performance is comparable to the MLG for fresh raw ground beef, fresh raw beef trim and beef carcass sampling sheets. For beef, a single enrichment protocol was validated for the synchronous detection of the STECs and Salmonella. 

The SIMUL-qPCR Listeria Species and Monocytogenes Assay were approved and assigned PTM Certification No. 062001. This validation study demonstrated that the performance of the SIMUL-qPCR Listeria Species and Monocytogenes Assay is comparable to the MLG, for frankfurters, RTE sliced turkey, and cooked eggs, and the BAM for chicken salad, ice cream, pasteurized milk, frozen shrimp, and stainless steel, plastic, rubber, ceramic tile and sealed concrete environmental surface sponges/swabs. 

Commenting on these approvals, Laura Cowin, Director of Technical Services at Applied Food Diagnostics, said: "Food laboratories face operational challenges by managing multiple protocols and detection platforms for pathogens."

"Providing a fully simultaneous multiplex SIMUL-qPCR platform where pathogens can be concurrently run on  the same platform is a significant milestone for those seeking operational efficiency.”

“We’re proud to offer the SIMUL-qPCR test kits, as we know there are food manufacturers who want to save time by consolidating run times.”

 “Overall sample management is enhanced with a single enrichment for both the STECs and Salmonella in beef matrices and the multiplexing of Listeria spp. and Listeria monocytogenes in one test,” said Cowin.