Total Collaborative Study Protocol_Solus One Salmonella v1 1

( a )  Dehydrated Buffered Peptone Water (BPW) Medium according to ISO 6579, CAT No. MED017 .  1  ( b )       Dehydrated Solus modified Buffered Peptone Water (Solus mBPW), CAT No. MED038 .  2  ( c )  Solus One Salmonella Supplement , CAT No. SALSUPP‐22.5 & SALSUPP‐112 .  3  ( d )  Dynex Technologies DS2 Instrument (Dynex DS2) . — Dynex Technologies, Chantilly, VA.  4  ( e )  Computer with DS2 Software .  5  ( f ) Dynex Technologies 300 µL Sample Tips .  6  ( g ) Dynex Technologies 1300 µL Reagent Tips .  7  ( h )  Sample Tubes .— 12 mm external diameter and 75 mm height.  8  ( i ) Polypropylene Tubes .— 2 mL and 15 mL and 25 mL.  9  ( j ) Frit filters .— Optional for matrices with a lot of particulate , CAT No. FRI001.  10  ( k ) 70% v:v Ethanol .— For preparation of the Solus One  Salmonella Supplement.  11  ( l ) Deionized water . 12  ( m )  Sterile collection sponge for sampling environmental surfaces .— World Bioproducts EZ Reach  13  Sponge Sampler with Hi Cap Neutralizing Buffer, or equivalent. 14  ( n )  Sterile collection swab for sample environmental surfaces . 15  ( o )  Letheen Broth neutralizing buffer .

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E. General Preparation 

( a ) Use aseptic techniques. 

( b ) Use filter laboratory bags during enrichment to minimize particulates. 

( c )  Separate work areas for the following: media preparation, sample preparation, and pathogen 

detection. 

( d ) Clean the work stations with a disinfectant of choice before and after use. (Sodium 

24  hypochlorite solution, phenol solution, Quaternary ammonium solution, etc.)  25  ( e ) Do not reuse kit disposables.  26  ( g )  Change pipette tips in between samples. 

Pathogen Detection (Manual Method) 

( a ) Equilibrate test kit to room temperature (15–25°C) an hour before use. 

( b ) Ensure that the bench processing time of inoculated samples is kept to a minimum, to avoid 

extensive growth of competing non‐ Salmonella bacteria. 

( c )  Start heating block or heat water bath to 85–100°C prior to initiating testing.  ( d ) Allow heated samples to cool to room temperature (15–25°C). This may be accelerated by 

placing the test tubes in cold water for 5 min. 

( e ) Where appropriate frit filters will be used with denatured matrices that contain high 

37  concentrations of particulates thereby minimizing sample handling errors.  38  ( f )  Change pipette tips in between samples. 

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Pathogen Detection (Automated Sample Preparation Method) 

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