Total Collaborative Study Protocol_Solus One Salmonella v1 1

supplement (2.22 mL Solus One  Salmonella  supplement per 1.0 L Solus mBPW Medium), were added to 

25 g and 375 g food test portions respectively. All food test samples were subsequently incubated for 

20–24 h at 41.5 ± 1°C. All food test samples were randomized, blind coded and then analyzed by the 

Solus One  Salmonella automated sample preparation method. 

All enriched test samples analyzed by the Solus One  Salmonella method, regardless of presumptive 

result, were culturally confirmed as described in FDA/BAM Ch. 5, Sections D (Isolation of  Salmonella ) 

through Section E.9 (1).  In addition, all enriched test samples were also confirmed using an alternative 

confirmation procedure by directly streaking the primary enrichments to either Bismuth Sulfite, Hektoen 

Enteric or Xylose Lysine Deoxycholate agars. Final confirmation was conducted using the Bruker MALDI 

Biotyper biochemical identification method, AOAC OMA 2017.09  (9).  

For the matrix study evaluation of the reference FDA/BAM Ch. 5 method, the 25 ± 2.5 g flavored 

ranch seasoning, honey mustard onion seasoning, paprika powder and black peppercorn portions were 

enriched with 225 ± 2.25 mL of Tryptic Soy Broth (TSB) and homogenized for 2 min by blending. For 

cinnamon powder, the 25 ± 2.5 g portions were enriched with 2,475 ± 24.75 mL of TSB and 

homogenized by hand massaging for 2 min. Following homogenization, test portions were allowed to 

stand for 60 ± 5 min at room temperature (24 ± 2°C), and if required, the pH of all matrix enrichments 

was re‐adjusted to 6.8 ± 0.2 before being incubated for 22–26 h at 35 ± 2°C. Following incubation, 0.1 ± 

0.02 mL and 1.0 ± 0.10 mL of each primary enriched sample was transferred into 10 mL of Rappaport 

Vassiliadis (RV) Broth and 10 mL of tetrathionate (TT) Broth respectively. RV broths were incubated in a 

circulating water bath for 22–26 h at 42 ± 0.2°C. As these food matrices contained high microbial 

backgrounds (>10 4  CFU/g), TT broths were incubated in a circulating water bath for 22–26 h at 43 ± 

0.2°C. The secondary enriched samples were culturally confirmed as described in the reference method 

FDA/BAM Ch. 5, Section D through E.9 (1); before final confirmation was conducted using the Bruker

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