AOAC-RI ERP Book MICRO.pdf

Page

5) When the run is complete, click “Save” and remove the samples from the

instrument.

Interpretation and Test Result Report

(a)

Viewing results

1) Software Version 2.3.1 (Build 49)

i. When the run is complete, select “Analysis” located on the top of

the main page of the software. In the “Analysis” box, import the

analysis settings into each channel by activating the “Quantitation

Analysis” window for the channel and select the respective file from

your directory with the “Import” function. Select “Ignore First” and

ignore the first 10 cycles for all four channels. For all channels

(Green, Crimson, Yellow, and Orange) select “Take Off Point

Adjustment”. Adjust the settings so that if the take off point was

calculated before cycle 15, then cycle 20 is used as the take of

point, then select “OK”. Set the threshold for channels Green,

Crimson, and Yellow to 0.035 and set the threshold for channel

Orange to 0.08.

(b)

Printing and Exporting Results

To export the results to Excel, go to the “File” menu, followed by “Save As” and

then “Excel Analysis Sheet”. The results will be saved in a .csv format. The

results may be saved to the notebook computer or an external flash drive. The

results may be printed by opening the “Report Browser” window, choosing a

report, and select “Show.” Choose “Print” on the report window.

(c)

Interpreting the results

Determining the presence or absence of pathogen DNA is carried out based on

the amplification of the target sequence and is visualized in real time on the

amplification plot generated by the application software of the real-time PCR

instrument used. A positive result is visible as a final point on the fluorescence

curve that lies clearly above the threshold.

Partial inhibition of the PCR due to the presence of detectable but tolerable

concentrations of inhibitors in the samples is typically indicated by a shift of the

internal control to higher C

values. As a guideline, the uninhibited internal

control should give a C

value ranging between 24 and 30. A cycle threshold

above 30 indicates inhibition.

In the event of a PCR inhibited internal control channel and a positive target

result, repeating the test is not necessary.

Confirmation

OMAMAN-36 E/ AOAC PTM Report

ERP Use Only

January 2017

AOAC Research Institute

Expert Review Panel Use O ly