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. 

( f )  Incubate the plate (containing the strips) at 37 ± 1°C for 30 ± 5 min.  ( g )  Following incubation, aspirate the contents of the wells, removing as much of the liquid as 

possible. Wash the wells 5–7 times with Washing Buffer ensuring completed filling and emptying of the 

wells through each wash cycle. Note: The washing technique is critical to assay performance; hence it is 

recommended to use a microplate washer.   

( h ) After completion of the washing steps, add 0.1 mL of Conjugate to each well, except for the  blank. Upon completion of the addition of the Conjugate, incubate the plate (containing the strips) at 37 

± 1°C for 30 ± 5 min. 

( i ) 

Repeat the aspiration and wash cycles. 

( j )  After completion of the second aspiration and washing steps, add 0.1 mL of TMB Substrate to 

each well, including the blank. Upon completion of the addition of the TMB Substrate, incubate the 

plate (containing the strips) for 30 ± 5 min at room temperature (15–25°C). 

( k )  After incubation, stop the reaction by adding 0.1 mL of Stop Solution to all wells, including the  “blank” well. Note: The Stop Solution will cause any blue color in wells to change to yellow.  ( l )  Where applicable read the optical densities within 10 min in a microplate reader using a 450  nm filter (do not use reference filter), or on the Dynex DS2 instrument using the “Plate Read Only” 

setting. Inspect the wells before reading for air bubbles and if present burst with a sterile needle. Zero 

the reader against the “blank” well before the other wells are read. 

Solus One Salmonella (Automated Sample Preparation Method) 

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

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