Total Collaborative Study Protocol_Solus One Salmonella v1 1

over at the end of a test strip the Positive or Negative Controls may be repeated.  1  ( f )  Incubate the plate (containing the strips) at 37 ± 1°C for 30 ± 5 min.  2  ( g )  Following incubation, aspirate the contents of the wells, removing as much of the liquid as  3  possible. Wash the wells 5–7 times with Washing Buffer ensuring completed filling and emptying of the  4  wells through each wash cycle. Note: The washing technique is critical to assay performance; hence it is  5  recommended to use a microplate washer.    6  ( h ) After completion of the washing steps, add 0.1 mL of Conjugate to each well, except for the  7  blank. Upon completion of the addition of the Conjugate, incubate the plate (containing the strips) at 37  8  ± 1°C for 30 ± 5 min.  9  ( i )  Repeat the aspiration and wash cycles.  10  ( j )  After completion of the second aspiration and washing steps, add 0.1 mL of TMB Substrate to  11  each well, including the blank. Upon completion of the addition of the TMB Substrate, incubate the  12  plate (containing the strips) for 30 ± 5 min at room temperature (15–25°C).  13  ( k )  After incubation, stop the reaction by adding 0.1 mL of Stop Solution to all wells, including the  14  “blank” well. Note: The Stop Solution will cause any blue color in wells to change to yellow.  15  ( l )  Where applicable read the optical densities within 10 min in a microplate reader using a 450  16  nm filter (do not use reference filter), or on the Dynex DS2 instrument using the “Plate Read Only”  17  setting. Inspect the wells before reading for air bubbles and if present burst with a sterile needle. Zero  18  the reader against the “blank” well before the other wells are read.  19  20  H. Solus One Salmonella (Automated Sample Preparation Method) 

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( a ) Prior to analysis, bring the entire contents of the kit to room temperature (15–25°C) by placing 

the kit on a sanitized bench top for 1 h prior to use. 

( b ) Determine the number of wells required for the test. Take the required number of strips from  25  the pouch and fit them to the frame provided. Note: Unused strips should be returned to the pouch and  26  stored at 2–8°C. 

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I. Analysis 

( a )  Prepare equipment

1) Turn on power to the Dynex DS2 instrument, followed by the power to the computer.  Open the Matrix software on the computer and enter login information. The Dynex DS2 

instrument will perform a self‐diagnostic check. 

( b )  Create a Run File 

1) In the DS‐ Matrix software, select “File”, “Worklist Editor”. Then select “New Sample 

Batch”. 

2) Enter your sample batch ID as required.  3) Enter the total number of samples in the batch. 

4) Sample ID may then be typed into the empty positions in the rack or assigned 

automatically using an onboard barcode scanner. 

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5) If using an onboard barcode scanner slide the sample tube racks onto the machine and 

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