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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