AOAC RI ERP Final Action Recommendations eBook

Collaborative Study Approved Protocol Expert Review Panel Use Only

preenrichment broth (UPB) (35°C for 22-26 h) for the FDA BAM method. After primary enrichment, ALL test portions (BAX and FDA BAM) will continue to selective enrichment. One mL of each enriched portion will be transferred to 10 mL TT broth and 0.1mL will be transferred to 10 mL RV broth. Note: BAM Method RV medium must be made from individual ingredients according to BAM formulation. Do not use commercial formulations. RV tubes will be incubated at 42 ± 0.2°C for 22-26 h using a circulating, thermostatically controlled water bath. TT tubes will be incubated at 35 ± 2°C for 22-26 h. Secondary enrichments will be streaked to bismuth sulfite (BS), xylose lysine desoxycholate (XLD), and Hektoen enteric (HE) agar plates and incubated at 35°C for 22-26 h. Isolated colonies will transferred to TSI and LIA slants and incubated 35 ± 2°C for 22-26 h. Salmonella colonies will be confirmed using serological (Somatic O and poly H agglutination) and biochemical procedures according to MLG Ch. 4.05. 13 14 Each level of each matrix will be analyzed and reported separately. Data may be excluded due to an 15 assignable cause, provided there is sufficient justification. Such data will not be included in the 16 statistical analysis. All data, even if excluded from the statistical analysis, will be reported (see Appendix 17 X-B of the AOAC Guidelines for raw data table format). 20 21 The statistical analysis will be performed in accordance with the AOAC Guidelines ( Appendix X-F ) using 22 the Least Cost Formulations, Ltd., AOAC Binary Data Interlaboratory Study Workbook (7). 1 2 3 4 5 6 7 8 9 10 11 12 6.0 Reporting Raw Data 18 19 7.0 Statistical Analysis

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7.1 7.2

Estimate the repeatability standard deviation (s r

) for qualitative methods.

Estimate the reproducibility (s R

) by calculating the standard deviation of the laboratory

POD values (s POD

) and associated 95% confidence intervals.

7.3 Estimate the difference of cross-laboratory POD (dLPOD) and associated 95% confidence intervals. If the confidence interval of a dLPOD does not contain a zero, then the

difference is statistically significant.

7.3.1 dLPODc is the difference between the candidate and reference LPOD values. 7.3.2 dLPOD is the difference between the presumptive and confirmed LPOD values. 32 33 In addition, the data will be analyzed by the Relative Limit of Detection (RLOD), as described in the most 34 current version of ISO 16140 (8). 37 38 BAX® System Real-Time PCR Assay for Salmonella Method for Detection of Salmonella in Foods and 39 Selected Environmental Surfaces 35 36 8.0 BAX® System Real-Time PCR Assay for Salmonella – Method

40 41

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