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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.
ESL Bldg. 400
1400 Harrison Avenue
PO Box 80400
Cincinnati, Ohio 45214
Route 141 & Henry Clay Rd.
Phone : 513-471-1300
Wilmington, DE 19880
Fax : 513-471-5600
Phone: 302-695-5473
Email:
pbird@qlaboratories.comFax : 302-695-5877
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
Collaborative Study Approved Protocol
Expert Review Panel Use Only