AOAC RI ERP Final Action Recommendations eBook

Dupont BAX Salmonella Pre- Collaborative Study For Expert Review Panel Use Only Do Not Distribute

The candidate presumptive and confirmatory results were compared by determining the difference 1 between the POD values for each matrix and concentration (dPOD CP = POD CP - POD CC ). If the confidence 2 interval of a dPOD does not contain zero, then the difference is statistically significant at the 5% level. 3 The candidate and reference method results were compared by determining the difference in POD 4 values between the candidate and reference methods for each matrix and concentration (dPOD C = POD C 5 - POD R ). If the confidence interval of a dPOD does not contain zero, then the difference is statistically 6 significant at the 5% level. 7 8 DuPont™ BAX® System Real-Time PCR Assay for Salmonella - Method for Detection of Salmonella in 9 Foods and Selected Environmental Surfaces 10 (Applicable to the detection of Salmonella in raw ground beef*, ground beef with soy, beef trim, 11 frankfurters, shrimp, ground turkey, chicken wings, poultry rinse*, dried eggs, shell eggs, fresh bagged 12 lettuce*, frozen peas, orange juice, cream cheese*, nonfat dry milk, ice cream, peanut butter, cocoa, 13 white pepper, infant formula, and dry pet food*, and on stainless steel*, ceramic tile, and plastic 14 surfaces) 15 * Previously validated as Performance Tested Method SM #081201 16 Safety Precautions 17 Kits – The reagents used in the BAX® System should pose no hazards when used as directed. Dispose of 18 lysate, PCR mixture and other waste according to your site practices. 19 Cycler/detector – Only qualified laboratory personnel should operate the cycler/detector. Do not 20 attempt to repair the instrument. Live power may still be available inside the unit even when a fuse has 21 blown or been removed. Refer to the BAX® System User Guide for maintenance procedures when 22 cleaning the unit or changing a fuse. The heating block can become hot enough during normal operation 23 to cause burns or cause liquids to boil. Wear safety glasses or other eye protection at all times during 24 operation. 25 Enrichment Broths – All enrichment broths may contain varying pathogens whether they contain 26 Salmonella or not and thus should be sterilized and disposed of using proper procedures following any 27 culture-based confirmatory steps. 28 A. Principle 29 The BAX® System uses the Polymerase Chain Reaction (PCR) to amplify a specific fragment of bacterial 30 DNA, which is stable and unaffected by growth environment. The fragment is a genetic sequence that is 31 unique to the genus Salmonella , thus providing a highly reliable indicator that the organism is present. 32 The BAX® System simplifies the PCR process by combining the requisite primers, polymerase and 33 nucleotides into a stable, dry, manufactured tablet already packaged inside the PCR tubes. After 34 amplification, these tubes remain sealed for the detection phase, thus significantly reducing the 35 potential for contamination with one or more molecules of amplified PCR product. 36 AOAC Official Method ####

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