AOAC ERP MICRO AUGUST 2018

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1 2 3 4 5 6 7 8 9

The first parameter, heat lysis duration, was evaluated by analyzing replicates after 13 and 17 minutes of heat treatment heated to 100ºC in a dry bath incubator. The second parameter, volume of sample to lysis solution, was evaluated by analyzing replicates by transferring either 18 µL or 22 µL of sample into lysis solution. The third parameter, volume of lysis solution into reaction tubes, was evaluated by adding either 22 µL or 18 µL of lysis solution to the MDA reaction

tubes. Table B below summarizes each treatment combination.

Table B: Robustness Treatment Conditions

Lysis Heat Time (Minutes)

Volume of Enrichment to Lysis

Volume of Lysis to Assay

Treatment Combination

1 2 3 4 5 6 7 8

13 13 13 13 17 17 17 17

18 µL 18 µL 22 µL 22 µL 18 µL 18 µL 22 µL 22 µL

18 µL 22 µL 18 µL 22 µL 18 µL 22 µL 18 µL 22 µL

10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33

Results

For the robustness evaluation, there were no observed discrepancies for 7 of the 8 treatment combinations when changing the operational parameters of the enrichment lysis time, volume of enrichment to lysis solution, and volume of lysis solution to assay. A summary of results is

displayed in Table 25 of the Appendix.

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Treatment Combinations 1, 2, 3, 5, 6, 7, 8

For these 7 treatment combinations, there were 6 presumptive positives out of 10 replicates for each variation evaluated. For the 5 non-target test portions, there were 0 presumptive positives

out of 5 replicates.

For the low inoculation level, a POD value of 0.60 was obtained with a 95% confidence interval of (0.31, 0.83). All 5 non-target test portions were negative with a POD value of 0.00 with a 95%

confidence interval of (0.00,0.43).

Treatment Combination 4

There were 5 presumptive positives out of 10 replicates. For the 5 non-target test portions, there were 0 presumptive positives out of 5 replicates. For the low invocation level, a POD value of