Total Collaborative Study Protocol_Solus One Salmonella v1 1

automated sample preparation method. All shredded cheddar cheese samples were also analyzed by 

the Solus One  Salmonella manual method. 

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

result, were culturally confirmed as described in the appropriate reference method MLG 4.09, Section 

4.6 (Selective Enrichment and Plating) through Section 4.9; or as described in BAM Ch. 5, Sections D 

(Isolation of  Salmonella ) through Section E.9. In addition, all enriched test samples were also confirmed 

using an alternative confirmation procedure by directly streaking the primary enrichments to either XLD 

or XLT4 (Xylose Lactose Tergitol) agars depending on matrix type. Final confirmation was conducted 

using the Bruker MALDI Biotyper biochemical identification method, AOAC OMA 2017.09  (8).  For the matrix study evaluation of the reference MLG 4.09 method, the 25 ± 2.5 g raw beef trim 

portions were enriched with 75 ± 1.5 mL of modified Tryptic Soy Broth, with the addition of casamino 

acids (mTSB+CAA). The samples were homogenized for 2 min and subsequently incubated for 15–24 h at 

42 ± 1°C. In contrast, the 100 ± 2 g pasteurized liquid egg portions were enriched with 900 ± 18 mL of 

BPW, homogenized for 2 min and subsequently incubated for 18–24 h at 35 ± 1°C. Following incubation, 

0.1 ± 0.02 mL and 0.5 ± 0.05 mL of each primary enriched sample was transferred into 10 mL of modified 

Rappaport Vassiliadis (mRV) Broth and 10 mL of Tetrathionate (TT) Hajna Broth respectively. The broths 

were incubated in a circulating water bath for 18–24 h at 42 ± 0.5°C. The secondary enriched samples 

were culturally confirmed as described in the reference method MLG 4.09, Section 4.6 through 4.9; 

before final confirmation was conducted using the Bruker MALDI Biotyper biochemical identification 

method, AOAC OMA 2017.09 . 

For the matrix study evaluation of the reference BAM Ch. 5 method, the 25 ± 2.5 g raw salmon fillet 

and shredded cheddar cheese portions were enriched with 225 ± 2.25 mL of Lactose Broth and 

homogenized for 2 min by blending. For bagged Romaine lettuce, the 25 ± 2.5 g portions were enriched

Page 31 of 47