Total Collaborative Study Protocol_Solus One Salmonella v1 1

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Ben Bastin (Study contact) 

Q Laboratories 

1930 Radcliff Drive   Cincinnati, Ohio 45204  

Email: bbastin@qlaboratories.com

2.0 8  9  A total of 14–16 collaborators will be solicited to participate in this study. A minimum of 10 valid  10  laboratory data sets are required to successfully complete the study. Each collaborator will receive  11  instructions for performing the study (Appendix 10.1) and required materials prior to the start of the  12  study. Training on the DS2 Instrument and the Solus One  Salmonella method will be provided as  13  necessary.      16  17  A collaborative study will be conducted in accordance with the AOAC INTERNATIONAL Methods  18  Committee Guidelines for Validation of Microbiological Methods for Food and Environmental Surfaces  19  (2). The Solus One  Salmonella method will be compared to the FDA BAM Chapter 5 reference method  20  for the detection of  Salmonella  in 375 g NFDM. A proprietary enrichment will be used for the Solus  21  method, so this study will be conducted as an unpaired study.    22  23  The NFDM will be screened for the presence of naturally contaminating  Salmonella and total aerobic  24  plate count ( note : for the FDA BAM method, if the level of background microflora is > 10 4  cfu/g, then the  25  tetrathionate (TT) broth must be incubated at 43 ± 0.2°C in a circulating, thermostatically‐controlled,  26  water bath, and if the level is ≤ 10 4  cfu/g, then TT should be incubated at 35 ± 2.0°C). Naturally  27  contaminated matrix is preferred if available. If naturally contaminated matrix is not available, artificially  28  contaminated matrix will be prepared. It is anticipated that artificial contamination will be required for  29  the NFDM. The matrix will be divided into 3 test materials. One test material remains non‐inoculated  30  and serves as the uncontaminated level, one test material is contaminated at a level that will produce a  31  reference method POD (POD R ) or candidate method POD (POD C ) in the range of 0.25–0.75  32  (approximately 0.2–2 CFU/test portion), and finally one test material is contaminated at a level to assure  33  a POD C of nearly 1.0 (approximately 10 CFU/test portion). Twelve replicate test portions per material  34  will be tested by the candidate method (36 test portions) and the reference method (36 test portions).    35  36  On the day that the analysis of the test samples is initiated, a 5‐tube 3‐level Most Probably Number  37  (MPN) estimation of contamination levels will be conducted using the FDA BAM reference method. The  38  MPN analysis scheme may also make use of the reference method replicates (see Appendix X‐A of the  39  AOAC Guidelines for details). The Least Cost Formulations, Ltd. (LCF) MPN Calculator‐Version 1.6 (3) will  40  be used to determine the MPN values and 95% confidence intervals. The MPN is reported as MPN/test  41  portion with 95% confidence intervals.  42 Collaborators  14  15  3.0 Study Design 

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