Total Collaborative Study Protocol_Solus One Salmonella v1 1

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( a ) Equilibrate test kit to room temperature (15–25°C) an hour before use.  ( b ) Start the Dynex DS2 and allow instrument to conduct the ‘Self‐Test’.  ( c )  Load required materials: pipette tips, reagents, washing buffer, etc. 

( d ) Empty waste. 

( e ) Start heating block or heat water to 85–100°C prior to initiating testing.  6  ( f )  Allow heated samples to cool to room temperature (15–25°C). This may be accelerated by  7  placing the test tubes in cold water for 5 min.  8  ( g )  Where appropriate use frit filters with denatured matrices that contain high concentrations of  9  particulates to minimize sample handling errors. 

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F. Sample Enrichment and Preparation

( a )  Raw Beef Trim (375 g test portions)

Pre‐warm 1,125 mL of Solus BPW (ISO) Medium supplemented with Solus  Salmonella  supplement  15  [4.44 mL Solus  Salmonella  supplement per 1,000 mL Solus BPW (ISO)] to 37 ± 1°C for at least 2 h prior to  16  its addition to a 375 g test portion in the sampling bag. Homogenize by hand massaging for 2 min ± 30 s  17  and incubate at 41.5 ± 1°C for 20–22 h.  18  ( b )  Pasteurized Liquid Egg (100 g test portions) 19  Pre‐warm 900 mL of Solus BPW (ISO) Medium supplemented with Solus  Salmonella  supplement  20  [4.44 mL Solus  Salmonella  supplement per 1,000 mL Solus BPW (ISO)] to 37 ± 1°C for at least 2 h prior to  21  its addition to a 100 g test portion in the sampling bag. Homogenize by hand massaging for 2 min ± 30 s  22  and incubate at 41.5 ± 1°C for 20–22 h.  23  ( c )  Raw Salmon (fillet), Cheddar Cheese (shredded) and Romaine Lettuce (bagged) (25 g test  24  portions) 25  Add 225 mL of Solus BPW (ISO) Medium supplemented with Solus  Salmonella  supplement [4.44 mL  26  Solus  Salmonella  supplement per 1,000 mL Solus BPW (ISO)] to a 25 g test portion in the sampling bag.  27  Homogenize for 2 min ± 30 s and incubate at 41.5 ± 1°C for 20–22 h.  28  ( d )  Paprika (powder) and Black Peppercorns (whole) (25 g test portions) 29  Add 225 mL of Solus mBPW Medium supplemented with Solus  Salmonella  supplement [2.22 mL  30  Solus  Salmonella  supplement per 1,000 mL Solus mBPW] to a 25 g test portion in the sampling bag.  31  Homogenize for 2 min ± 30 s and incubate at 41.5 ± 1°C for 20–24 h.  32  ( e )  Cinnamon (powder) (25 g test portions) 33  Add 1,225 mL of Solus mBPW Medium supplemented with Solus  Salmonella  supplement [2.22 mL  34  Solus  Salmonella  supplement per 1,000 mL Solus mBPW] to a 25 g test portion in the sampling bag.  35  Homogenize by hand massaging for 2 min ± 30 s and incubate at 41.5 ± 1°C for 20–24 h.  36  ( f )  Honey Mustard Onion Seasoning and Flavored Ranch Seasoning (375 g test portions) 37  Add 3,375 mL of Solus mBPW Medium supplemented with Solus  Salmonella  supplement [2.22 mL  38  Solus  Salmonella  supplement per 1,000 mL Solus mBPW] to a 375 g test portion in the sampling bag.  39  Homogenize by hand massaging for 2 min ± 30 s and incubate at 41.5 ± 1°C for 20–24 h.  40  ( g )  Non‐Fat Dry Milk Powder (375 g test portions) 41  Pre‐warm 1,125 mL of Solus BPW (ISO) Medium supplemented with Solus  Salmonella  supplement  42

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