AOAC RI ERP Final Action Recommendations eBook

Collaborative Study Approved Protocol Expert Review Panel Use Only

10.2 Instructions to Collaborators

Dear Collaborators, Thank you for your participation in the AOAC BAX ® System Real-Time PCR Assay for Salmonella collaborative study. This outline provides detailed information needed to conduct each method and steps to take when reporting results. We look forward to working with you on this project.

Sincerely,

Co-Study Director

Co-Study Director

Morgan Wallace, Ph.D.

Patrick Bird

DuPont Qualicon

Q Laboratories, Inc. 1400 Harrison Avenue Cincinnati, Ohio 45214 Phone : 513-471-1300 Fax : 513-471-5600

ESL Bldg. 400 PO Box 80400

Route 141 & Henry Clay Rd.

Wilmington, DE 19880 Phone: 302-695-5473

Email: pbird@qlaboratories.com

Fax : 302-695-5877 Email: Morgan.Wallace@usa.dupont.com

INTRODUCTION: Study Goals

The purpose of this collaborative study is to compare the performance of the BAX ® System Real-Time PCR Assay for Salmonella to the USDA/FSIS-MLG 4.05 and the FDA/BAM Chapter 5 reference methods for the detection of Salmonella species. This evaluation will be conducted in a multilaboratory study analyzing 2 foods: orange juice and hot dogs. Test portions from 3 different contamination levels, including un-inoculated negative controls, will be sent to collaborators who will perform the BAX ® System Real-Time PCR Assay for Salmonella along with the U.S. reference methods: USDA/FSIS Microbiology Laboratory Guidebook Chapter 4.05 and FDA/BAM Chapter 5. The following sample sizes will be tested: orange juice (25 gram sample size) and hot dogs (325 gram sample size)

Description of the BAX ® System Real-Time PCR Assay for Salmonella

The BAX® System Real-Time PCR Assay for Salmonella uses Polymerase Chain Reaction (PCR) to amplify a specific fragment of bacterial DNA, which is stable and unaffected by growth environment. The fragment is a genetic sequence that is unique to the genus Salmonella , thus providing a highly reliable indicator that the organism is present. The BAX® System simplifies the PCR process by combining the requisite primers, polymerase and nucleotides into a stable, dry, manufactured tablet already packaged inside the PCR tubes. After amplification, these tubes remain sealed for the detection phase, thus

26