AOAC RI ERP Final Action Recommendations - BOOK

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1
Follow prompts in the Rack Wizard to enter identifying data on the entire rack and on the individual 2
samples. 3
Place one cluster tube per sample in a cluster tube rack. Add 200 µL of prepared lysis reagent to 4
each tube (lysis tube). Transfer 5 µL of each pre-enriched sample to the corresponding lysis tube and 5
ensure the tubes are capped. Incubate lysis tubes at 37ºC for 20 minutes, then at 95ºC for 10 minutes in 6
either individual heating blocks or the DuPont Thermal Block. Cool lysates for at least five minutes in 7
either a refrigerated cooling block or the automated thermal block prior to the final transfer to PCR 8
tubes. 9
Warm up the cycler/detector by selecting RUN FULL PROCESS from the menu bar of the application 10
window. 11
Place a PCR tube holder on the PCR cooling block. Insert one PCR tube per sample into the holder 12
and remove caps. Using a multi-channel pipette, transfer 30 µL of sample lysate to a PCR tube. Cap PCR 13
with the flat optical cap strips provided in the kit. 14
Follow screen prompts to load samples into the cycler/detector and begin the program. At the 15
completion of the PCR and detection process, follow the screen prompts to remove samples and display 16
results. 17
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F. Interpretation of Test Result 19
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The results are recorded on the rack display or from a spreadsheet printout of the results 21
(called Detail View). Negative results are indicated by the green circle with (-) symbol, positive 22
results are indicated by the red circle with (+) symbol, and indeterminate results are indicated with 23
a yellow circle with (?) symbol. A yellow circle with a (?) symbol and a red slash indicated a low 24
signal or signal error. 25
BAX® System results were displayed as follows: 26
Green (-) Negative for Salmonella Yellow (?)
Indeterminate
result
Red (+) Positive for Salmonella
Yellow (?) with
red slash
Signal error
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G. Confirmation 29
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Presumptive positive samples must be confirmed culturally as described in AOAC Official Method 31
967.26 , AOAC Official Method 2000.06 (see ), the U.S. FDA Bacteriological 32
Analytical Manual (see 33
34
the USDA-FSIS Microbiology Laboratory Guidebook (see 35
, or the Health Canada Compendium of Analytical 36
Collaborative Study Approved Protocol
Expert Review Panel Use Only