Total Collaborative Study Protocol_Solus One Salmonella v1 1

Each collaborator will receive a complete set of test materials (uncontaminated, low, and high) for each  1  method (candidate and reference) and will also receive one known uncontaminated sample to  2  determine the aerobic plate count (4). All participating collaborators will initiate sample analysis and  3  aerobic plate count for each matrix on the same day. One additional control (negative) test portion will  4  be provided to each collaborator for each matrix to determine the total plate count on the day of  5  analysis.  5.1 Each collaborator will receive one set of 375 g test portions (12 uncontaminated, 12  low, 12 high) to run on the Solus One Salmonella method, and one set of 25 g test  portions to run on the FDA BAM method. Each set will be blind coded so that the  6  7  8  9  5.2 The 375 g set of test portions will be enriched in Solus One  Salmonella 1125 mL Solus  One enrichment broth (BPW + supplement) at 41.5 ± 1°C for 21–22 h. Collaborators who  have the Dynex DS2 instrument in their laboratories will conduct the Solus One  Salmonella method using the Dynex instrument (automated procedure). Collaborators  who do not have the Dynex DS2 instrument in their laboratories will conduct the Solus  One  Salmonella method following the manual procedure. It is anticipated that there will  5.3 The 25 g set of test portions will be enriched in 225 mL brilliant green water contained  in sterile 500 mL Erlenmeyer flask or another appropriate container for 24 ± 2 h at 35°C. 5.4 After primary enrichment, ALL test portions (Solus One  Salmonella and FDA BAM) will  continue to selective enrichment. One (1) mL of each enriched portion will be  transferred to 10 mL TT broth and 0.1 mL will be transferred to 10 mL RV broth. Note:  BAM Method RV medium must be made from individual ingredients according to BAM  formulation.  Do not use commercial formulations. RV tubes will be incubated at 42 ±  0.2°C for 22–26 h using a circulating, thermostatically controlled water bath. TT tubes  will be incubated at 35 ± 2°C for 22–26 h. Secondary enrichments will be streaked to  bismuth sulfite (BS), xylose lysine desoxycholate (XLD), and Hektoen enteric (HE) agar  plates and incubated at 35°C for 22‐26 h. Isolated colonies will be transferred to TSI and  LIA slants and incubated 35 ± 2°C for 22‐26 h. Salmonella  colonies will be confirmed  using serological (Somatic O and poly H agglutination) and biochemical procedures  according to FDA BAM Ch. 5. The recovered isolates will be identified by the API20E  (Official Method 978.24 ), VITEK2 GN (Official Method 2011.17 ) or Bruker MALDI (OMA  5.5 In addition, all Solus One  Salmonella enriched test portions will be confirmed using an  alternative confirmation procedure. Each portion will be streaked directly from the  primary enrichment onto selective agar, as described by BAM Ch. 5. Confirmation will  continue through to identification from the selective agar as described in 5.4.    be an even mix of collaborators (automated and manual).   2017.09 ) depending on availability.   contamination level is unknown to the collaborator.  

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Reporting Raw Data 

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