Table 2 |
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|
Intra- and inter-assay co-efficient of variation for EBNA-1 and BHRF-1 QPCRs. |
||||||
|
DNA Target (copies/5 ul) |
Mean CT |
Mean R-G 6000™ Results (copies/5ul) |
Standard Deviation of R-G 6000™ Results (copies/5ul) |
Mean % Variation |
COV (%) |
Mean R2 |
|
|
||||||
|
EBNA-1 Intra-Assay Variation (same day) |
||||||
|
|
||||||
|
100,000 |
18.03 |
87,329 |
6,670 |
12.68% |
7.64 |
0.991 |
|
10,000 |
21.28 |
11,735 |
3,092 |
26.30% |
26.34 |
|
|
1,000 |
25.21 |
1,057 |
100 |
7.00% |
9.50 |
|
|
100 |
29.10 |
103 |
38 |
32.12% |
37.25 |
|
|
10 |
32.91 |
11 |
6 |
46.16% |
57.50 |
|
|
|
||||||
|
EBNA-1 Intra-Assay Variation (different days) |
||||||
|
|
||||||
|
100,000 |
16.94 |
89,643 |
8,164 |
11.00% |
9.11 |
0.998 |
|
10,000 |
20.31 |
10,678 |
1,207 |
10.00% |
11.31 |
|
|
1,000 |
23.85 |
1,133 |
129 |
16.00% |
11.41 |
|
|
100 |
27.65 |
102 |
4 |
3.00% |
3.90 |
|
|
10 |
31.42 |
10 |
2 |
17.88% |
18.95 |
|
|
|
||||||
|
BHRF-1 Intra-Assay Variation (same day) |
||||||
|
|
||||||
|
100,000 |
17.23 |
97,884 |
9,144 |
8.08% |
9.34 |
0.994 |
|
10,000 |
20.91 |
9,852 |
542 |
4.45% |
5.50 |
|
|
1,000 |
24.38 |
1,146 |
202 |
16.12% |
17.64 |
|
|
100 |
28.43 |
94 |
19 |
17.70% |
20.51 |
|
|
10 |
31.95 |
11 |
5 |
35.38% |
41.91 |
|
|
|
||||||
|
BHRF-1 Intra-Assay Variation (different days) |
||||||
|
|
||||||
|
100,000 |
18.05 |
105,387 |
4,621 |
6.02% |
4.38 |
0.997 |
|
10,000 |
21.75 |
9,779 |
818 |
6.23% |
8.37 |
|
|
1,000 |
25.23 |
1,042 |
141 |
11.63% |
13.55 |
|
|
100 |
29.06 |
89 |
18 |
13.76% |
19.88 |
|
|
10 |
32.66 |
12 |
2 |
25.30 |
16.53 |
|
|
|
||||||
|
EBNA-1 Inter-Assay Variation |
||||||
|
|
||||||
|
100,000 |
19.87 |
101,644 |
14,058 |
10.99% |
28.35 |
0.990 |
|
10,000 |
23.75 |
10,660 |
1,471 |
13.65% |
13.80 |
|
|
1,000 |
27.68 |
1,084 |
191 |
18.67% |
17.60 |
|
|
100 |
31.71 |
111 |
49 |
33.58% |
43.75 |
|
|
10 |
35.86 |
12 |
9 |
65.03% |
75.85 |
|
|
|
||||||
|
BHRF-1 Inter-Assay Variation |
||||||
|
|
||||||
|
100,000 |
17.30 |
109,065 |
14,266 |
10.01% |
13.08 |
0.990 |
|
10,000 |
21.49 |
9,209 |
2,154 |
16.84% |
23.39 |
|
|
1,000 |
25.49 |
860 |
251 |
22.19% |
29.15 |
|
|
100 |
29.01 |
108 |
49 |
35.53% |
45.33 |
|
|
10 |
32.29 |
15 |
8 |
69.41% |
57.23 |
|
|
|
||||||
|
Abbreviations: CT, cycle threshold; Mean % variation, average percentage variation between the calculated (Rotor-Gene results) and the given concentration (DNA target); COV, co-efficient of variation is the ratio of standard deviation to the mean; R2-value, square root of the correlation co-efficient - in quantitation PCR describes the percentage of the data which matches the hypothesis that the standards conform to a line of best fit. |
||||||
|
Lay et al. Virology Journal 2010 7:252 doi:10.1186/1743-422X-7-252 |
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