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Activity 2 Figure 8 shows three examples of quantitative PCR results from three different patients. Use what you have learned about the negative control, positive control, and threshold of detection to label the lines. Notice the difference in the number of cycles (x axis) that each sample takes to peak in its fluorescence (y axis). Using your knowledge, interpret results of each test by indicating "positive," "negative," or "ambiguous" on the lines provided below. Patient #1 Result: Negative Patient #2 Result: Positive Patient #3 Result: Ambiguous Questions 11. What could contribute to ambiguous results from samples? Hint: Think about differences in the viral load (the total amount of virus inside a person) between people, differences in sample collection methods, the differences in sample preparation methods. 12. Suggest two factors that may contribute to an ambiguous test result. qPCR Report Fluorescence intensity 0.25 0.5 0.75 1.0 0 5 10 15 20 25 30 35 40 Number of PCR cycles Patient #1 sample qPCR Report Fluorescence intensity 0.25 0.5 0.75 1.0 0 5 10 15 20 25 30 35 40 Number of PCR cycles Patient #2 sample qPCR Report Fluorescence intensity 0.25 0.5 0.75 1.0 0 5 10 15 20 25 30 35 40 Number of PCR cycles Patient #3 sample Figure 8. RT-qPCR Reports for Three Different Patients

          Activity 2
Figure 8 shows three examples of quantitative PCR results from three different patients. Use what you have learned about the negative control, positive control, and threshold of detection to label the lines. Notice the difference in the number of cycles (x axis) that each sample takes to peak in its fluorescence (y axis). Using your knowledge, interpret results of each test by indicating "positive," "negative," or "ambiguous" on the lines provided below.

Patient #1 Result: Negative
Patient #2 Result: Positive
Patient #3 Result: Ambiguous

Questions
11. What could contribute to ambiguous results from samples? Hint: Think about differences in the viral load (the total amount of virus inside a person) between people, differences in sample collection methods, the differences in sample preparation methods.

12. Suggest two factors that may contribute to an ambiguous test result.

qPCR Report
Fluorescence intensity
0.25 0.5 0.75 1.0
0 5 10 15 20 25 30 35 40
Number of PCR cycles
Patient #1 sample

qPCR Report
Fluorescence intensity
0.25 0.5 0.75 1.0
0 5 10 15 20 25 30 35 40
Number of PCR cycles
Patient #2 sample

qPCR Report
Fluorescence intensity
0.25 0.5 0.75 1.0
0 5 10 15 20 25 30 35 40
Number of PCR cycles
Patient #3 sample

Figure 8. RT-qPCR Reports for Three Different Patients

Activity 2
Figure 8 shows three examples of quantitative PCR results from three different patients. Use what you have learned about the negative control, positive control, and threshold of detection to label the lines. Notice the difference in the number of cycles (x axis) that each sample takes to peak in its fluorescence (y axis). Using your knowledge, interpret results of each test by indicating "positive," "negative," or "ambiguous" on the lines provided below.

Patient #1 Result: Negative
Patient #2 Result: Positive
Patient #3 Result: Ambiguous

Questions
11. What could contribute to ambiguous results from samples? Hint: Think about differences in the viral load (the total amount of virus inside a person) between people, differences in sample collection methods, the differences in sample preparation methods.

12. Suggest two factors that may contribute to an ambiguous test result.

qPCR Report
Fluorescence intensity
0.25 0.5 0.75 1.0
0 5 10 15 20 25 30 35 40
Number of PCR cycles
Patient #1 sample

qPCR Report
Fluorescence intensity
0.25 0.5 0.75 1.0
0 5 10 15 20 25 30 35 40
Number of PCR cycles
Patient #2 sample

qPCR Report
Fluorescence intensity
0.25 0.5 0.75 1.0
0 5 10 15 20 25 30 35 40
Number of PCR cycles
Patient #3 sample

Figure 8. RT-qPCR Reports for Three Different Patients

Added by James V.

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