Review Book

General Comments about the Method Scope/Applicability:

GENE-UP Red page (manuscript page) 1. 8 (3)-2nd paragraph, line 4-should read “raw chicken breast” 2. 8(3)-raw fish is missing from the matrices list. 3. 9(4)-NCTC-spell out source and provide location (city, state) 4. 13(8)-missing are cooked ham, lettuce, liquid whole egg.

5. 15(10)-(g) should be Troemner Vortex-type mixer, and the Advanced digital Heavy should be (1) under that with 120V and 230V under that, then (2) should be Vortex-Genie Pulse, with (h) GENE-UP Lysis Rack Adaptor under it (see package insert) 6. 16(11)-CHROMID Salmonella is missing from page 150(2) list under Confirmation. 7. 17(12)-(g) “…cleaned and decontaminated” is also Safety. (also in IFU) 8. 17(12)-1st paragraph of Safety should be added to IFU. 9. 18(13)-D. Sample Enrichment-length of time for homogenization is missing here and in IFU; on page 10(5) it says the test portions were homogenized for 2 min. 10. 18(13)-(a)-raw ground beef (25 g) is not in AOAC RI Approved Protocols table in IFU 153(5). 11. 18(13)-(a)-IFU says BPW at room temperature, not in manuscript. Also, need to define “room temperature” 12. 19(14)-(b)-milk chocolate (25 g) missing in IFU 13. 19(14)-raw ground beef (375 g) is missing here and in table in IFU 14. 19(14)-(k)-garlic powder-temperature of incubation is 35 degrees in IFU instead of 42 degrees in manuscript. 15. 19(14)-(l)-IFU states 1125 mL for creamy peanut butter instead of 3375 shown here. 16. 19(14)-(m)-Alternative method for dark chocolate not in IFU. 17. 19(14)-(n)-milk chocolate (375 g) not in IFU. 18. 20(15)-(p)-environmental samples not in IFU (except in note). 19. 20(15)-note re alternative method-“Extending the enrichment time to 24 hours allows for performance improvement of the alternative method only for large portions of raw meat products.” Was this in previous validations? This should be referred to since that data is not shown in this study. 208. 24(19)-I-add “as appropriate for the matrix” to the sentence. 21. 24(19)-I(a)CHROMID Salmonella is not in IFU (Confirmation of Positive Results Obtained using AOAC RI Approved Protocols) 22. 24(19)-I(b)-VITEK GN and VITEK MS are not in IFU 23. 25(20)-I(g)-ViTEK GN and VITEK MS are not in IFU 24. 25(20)-J(b)-IFU missing +/- 1 degrees 25. 27(22) and 28(23)-It would be good to describe Alternative Confirmation vs. Traditional Confirmation in these sections as a reminder. 26. 29-30(24-25)-Was the benefit of using the alternative method demonstrated, or why a laboratory would use it instead of traditional confirmation method?

Pros/Strengths of the Manuscript:

Participating laboratories pointed out its ease of use compared to the reference method with high background microflora.

Cons/Weaknesses of the Manuscript:

None noted

Is the Validation Study Manuscript in a format acceptable to AOAC?

Yes