Table 3.
Summary of the fluorescence ratio based method and the Ct base method for quantification of predefined standards
| Group No. In predefined standards | Predefined allele ratio YMDD: | Kf method | Baseline subtracted fluorescence ratio | SDM | ||||||
| YIDD | Mean (n = 4)a | S.D (n = 4)b | Intra C.Vc | Mean (n = 4)d | S.D (n = 4)e | Intra C.Vc | Mean (n = 4)f | S.D (n = 4)g | Intra C.Vc | |
| 1 | 90%: 10% | 2.68 | 0.32 | 11.80% | 8.31 | 0.99 | 11.95% | 6.46 | 3.06 | 47.29% |
| 2 | 80%: 20% | 1.40 | 0.08 | 5.45% | 4.25 | 0.27 | 6.45% | 2.73 | 0.09 | 3.30% |
| 3 | 70%: 30% | 0.80 | 0.14 | 17.22% | 2.15 | 0.69 | 31.87% | 1.46 | 0.14 | 9.32% |
| 4 | 60%: 40% | 0.54 | 0.03 | 6.11% | 1.38 | 0.17 | 12.22% | 1.09 | 0.27 | 24.43% |
| 5 | 50%: 50% | 0.40 | 0.03 | 7.30% | 1.12 | 0.17 | 15.59% | 0.65 | 0.07 | 10.38% |
| 6 | 40%: 60% | 0.30 | 0.02 | 7.77% | 0.76 | 0.10 | 13.09% | 0.51 | 0.04 | 8.56% |
| 7 | 30%: 70% | 0.22 | 0.00 | 1.91% | 0.62 | 0.03 | 4.89% | 0.32 | 0.08 | 25.12% |
| 8 | 20%: 80% | 0.15 | 0.01 | 3.67% | 0.39 | 0.03 | 6.57% | 0.43 | 0.18 | 42.29% |
| 9 | 10%: 90% | 0.07 | 0.01 | 10.24% | 0.17 | 0.01 | 5.98% | 0.01 | 0.02 | 113.30% |
| Av. | Average CV | 7.94% | 12.07% | 31.56% | ||||||
The spreadsheet used for this analysis is available as Supplementary Material.1 [see Additional file 1]
a. Arithmetic mean Kf of 4 replicates
b. Combined Standard deviation Kf for 4 replicates
c. (SD/mean) × 100%
d. Fluorescence ratio arithmetic mean of 4 replicates
e. Combined Standard deviation fluorescence ratio for 4 replicates
f. Arithmetic mean 2-ΔCt of 4 replicates
g. Standard deviation of 2-ΔCt for 4 replicates