Total Collaborative Study Protocol_Solus One Salmonella v1 1

1.0 1  2  The purpose of this collaborative study is to determine the performance of the candidate method  3  among collaborators for the detection of  Salmonella . Estimates of repeatability, reproducibility and  4  probability of detection (POD) will be evaluated. Collaborators will analyze one matrix at 3  5  contamination levels comparing the performance of the candidate method to appropriate reference  6  culture method. The Solus One  Salmonella  is the candidate method. The U.S. Food and Drug  7  Administration Bacteriological Analytical Manual  (FDA BAM) Chapter 5 (1) is the reference method for  8  this study. 9  10  1.1 Solus One  Salmonella Introduction  The Solus One  Salmonella method is an en zyme‐linked immunosorbent assay ( ELISA) designed  to detect  Salmonella  ( Salmonella enterica  subspecies enterica ,  salamae , arizonae , diarizonae ,  houtenae ,  indica and  Salmonella bongori ) in select foods and environmental samples following  enrichment containing a proprietary supplement. The method utilizes antibodies bound to the  wells of microplate strips that are specific to  Salmonella antigens and detects both motile and  non‐motile  Salmonella . Heat inactivated/lysed samples are added to the wells of the plate and  sample analysis is conducted following a manual or a fully automated protocol on the Dynex DS2  instrument. Results are based on optical density levels resulting from a color change reaction in  the wells. Presumptive positive or negative results can be obtained in as little as 22 h for most 

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food samples and 18 h for environmental samples. See Attachments 1 and 2. 

Summary of the Performance Tested Method SM  Validation 

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The Solus One  Salmonella method was certified by the AOAC Research Institute in October,  2018 and designated Performance Tested Method SM  (PTM) 101801 for six food matrices; raw  beef trim (375 g), non‐fat dry milk powder (NFDM, 375 g), pasteurized liquid egg (100 g), raw  salmon (25 g), cheddar cheese (25 g), Romaine lettuce (25 g), and two environmental surfaces  (stainless steel and polystyrene). A method modification was approved in August 2019 to  include herbs, spices and flavorings; whole black peppercorns, paprika powder and cinnamon  powder (25 g sample size) plus honey mustard onion seasoning and flavored ranch seasoning 

(375 g sample size). See Attachments 3 and 4. 

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Study Directors 

Ray Wakefield 

Solus Scientific Solutions, Ltd. 

9 Mansfield Networkcentre, Millennium Business Park 

Concorde Way, Mansfield 

Nottinghamshire NG19 7JZ, United Kingdom  Email:  ray.wakefield@solusscientific.com

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