Total Collaborative Study Protocol_Solus One Salmonella v1 1

Dynex DS2 .— Improper use of the Dynex DS2 may cause personal injury or damage to the instrument.  1  For safe use, the Dynex DS2 must be operated only by qualified laboratory personnel who have been  2  appropriately trained. 3  Enrichment .— Salmonella  is a Biosafety Level 2 organism. Biological samples such as enrichments have  4  the potential to transmit infectious diseases. Follow all applicable local, state/provincial, and/or national  5  regulations on disposal of biological wastes. Wear appropriate protective equipment which includes but  6  is not limited to: protective eyewear, face shield, clothing/laboratory coat, and gloves. All work should  7  be conducted in properly equipped facilities utilizing the appropriate safety equipment (e.g., physical  8  contaminant devices). Individuals should be trained in accordance with applicable regulatory and  9  company/institution requirements before working with potentially infectious materials. All enrichment  10  broths should be sterilized following any culture based confirmatory steps through heat denaturation by  11  autoclaving at 121°C for 15 min.  16  proprietary media supplement – Solus One  Salmonella  supplement; for the rapid and specific detection  17  of  Salmonella  species in select foods and environmental samples. The modification described in this  18  report details the addition of five matrices: honey mustard onion seasoning and flavored ranch  19  seasoning (375 g sample size) plus cinnamon powder, paprika powder and whole black peppercorns (25  20  g sample size) in combination with a proprietary media – Solus modified Buffered Peptone Water  21  (mBPW).  22  Solus One  Salmonella  relies on antibodies attached to the wells of microplate strips by non‐covalent  23  biological interactions that are highly specific to Salmonella antigens. Samples are heat treated and an  24  aliquot is added to the antibody coated wells.  25  Salmonella  specific antigens present in the samples will bind immunologically to the antibody. After  26  washing to remove unbound material, an enzyme‐labelled antibody will bind to the captured proteins  27  and thus to the well. After a second wash step to remove any unbound enzyme‐antibody, the enzyme  28  substrate is added. The substrate reacts in the presence of the enzyme producing a blue color change in  29  the sample well. The substrate reaction is stopped after 30 minutes with the addition of dilute sulfuric  30  acid changing any blue color present in the wells to yellow (3). Optical densities resulting from this color  31  change are read within 10 minutes in a generic plate reader using a 450 nm filter (e.g. a microplate  32  reader or a Dynex DS2 instrument plate reader), where a result of an OD 450 < 0.200 is considered to be  33  negative for the target pathogen and OD 450 ≥ 0.200 is considered to be positive for the target pathogen.  12  13  14  15  A. Principle  Solus One  Salmonella  is an antibody‐based high sensitivity ELISA method paired with media and our 

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B. Apparatus 

( a )  Refrigerator .— capable of maintaining 2–8°C. 

( b )  Incubators .— capable of maintaining 37 ± 1°C and 41.5 ± 1°C.  ( c )  Circulating water bath .— capable of maintaining 42–50°C.  ( d )  Heating block or water bath .— capable of maintaining 85–100°C. 

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