226 / 453
14
72 count strip tubes for the manual preparation method. Ensure that the
1
locking ring is in place and select “Next”. Verify that the reaction volume
2
is 20 µL and select “Next”. Select “Next” after verifying the temperature
3
profile. Select “Start Run” to start the Rotor-Gene Q instrument.
4
5)
When the run is complete, click “Save” and remove the samples from the
5
instrument.
6
7
G.
Interpretation and Test Result Report
8
(a)
Viewing results
9
10
1)
Software Version 2.3.1 (Build 49)
11
12
i.
When the run is complete, select “Analysis” located on the top of the main
13
page of the software. In the “Analysis” box, import the analysis settings to
14
each channel by activating the “Quantitation Analysis” window for the
15
channel and select the respective file from your directory with the
16
“Import” function. Select “Ignore First” and ignore the first 10 cycles for
17
all four channels. For all channels (Green, Crimson, Yellow, and Orange)
18
select “Take Off Point Adjustment”. Adjust the settings so that if the take
19
off point was calculated before cycle 15, then cycle 20 is used as the take
20
of point, then select “OK”. Set the threshold for channels Green, Crimson,
21
and Yellow to 0.035 and set the threshold for channel Orange to 0.08.
22
23
(b)
Printing and Exporting Results
24
25
To export the results to Excel, go to the “File” menu, followed by “Save As” and then
26
“Excel Analysis Sheet”. The results will be saved in a .csv format. The results may be
27
saved to the notebook computer or an external flash drive. The results may be printed
28
by opening the “Report Browser” window, choosing a report, and select “Show.”
29
Choose “Print” on the report window.
30
Results of Collaborative Study
31
32
This collaborative study involved a method comparison evaluation of the
mericon
E. coliO157
33
Screen Plus and
mericon
E. coliSTEC O-Type Pathogen Detection Assays to the USDA FSIS
34
MLG 5.09 reference method for raw ground beef. A total of five laboratories throughout the
35
continental United States participated in this study. Four of the laboratories had 3 separate
36
analysts participate and the fifth laboratory had one participant. Twelve of the 13 total
37
participants submitted data. One participant, identified as Laboratory #2, did not incubate their
38
test portions for the correct duration. When the error was determined, no further sample analysis
39
was conducted and no data was submitted. Each participant analyzed 36 paired test portions for
40
each assay: 12 inoculated with a high level of
E. coli
O157:H7, 12 inoculated with a low level of
41
E. coli
O157:H7, and 12 un-inoculated controls.
42
43
OMAMAN-36 A : Collaborative Study Manuscript
For ERP Use Only
January 2017
AOAC Research Institute
Expert Review Panel Use Only