Table 4. Comparative multiplex TaqMan qPCR sensitivity assays value.
| Template conc. per reaction | Ct(SD) value of corresponding channel using Rotor-Gene Multiplex qPCR kit | Ct(SD) value of corresponding channel using SsoFast Probe Supermix | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Orange | Yellow | Red | Crimson | Green | Orange | Yellow | Red | Crimson | Green | ||
| R2 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | 0.996 | 0.995 | 0.996 | 0.996 | 0.997 | |
| Y | -3.31 | -3.29 | -3.37 | -3.37 | -3.35 | -3.85 | -4.30 | -3.83 | -3.85 | -3.85 | |
| Ex | 1.01 | 1.01 | 0.98 | 0.98 | 0.99 | 0.82 | 0.71 | 0.83 | 0.82 | 0.82 | |
| *Ct(SD) values | 1 ng | 9.32(0.36) | 9.68(0.36) | 11.71(0.37) | 9.76(0.32) | 12.66(0.43) | 9.13(0.36) | 8.56(0.43) | 11.93(0.29) | 10.41(0.31) | 14.20(0.41) |
| 100 pg | 12.52(0.29) | 12.84(0.28) | 14.92(0.39) | 12.97(0.35) | 15.73(0.37) | 12.11(0.11) | 11.63(0.18) | 14.78(0.17) | 13.33(0.15) | 17.46(0.11) | |
| 10 pg | 15.70(0.30) | 16.00(0.34) | 18.19(0.31) | 16.21(0.35) | 18.91(0.50) | 16.28(0.16) | 15.70(0.11) | 19.28(0.19) | 17.55(0.03) | 21.58(0.11) | |
| 1 pg | 19.17(0.13) | 19.39(0.12) | 21.74(0.17) | 19.75(0.19) | 22.55(0.13) | 20.89(0.56) | 21.12(0.71) | 23.30(0.76) | 21.97(0.53) | 25.86(0.60) | |
| 100 fg | 22.42(0.12) | 22.65(0.13) | 24.98(0.17) | 23.07(0.14) | 25.73(0.16) | 24.74(0.11) | 25.59(0.17) | 27.26(0.15) | 25.90(0.07) | 29.68(0.15) | |
| 10 fg | 25.84(0.21) | 26.09(0.17) | 28.48(0.17) | 26.59(0.20) | 29.23(0.20) | 28.34(0.40) | 29.80(0.59) | 31.03(0.48) | 29.57(0.50) | 33.47(0.50) | |
| 1 fg | 29.09(0.10) | 29.36(0.08) | 31.83(0.08) | 29.87(0.02) | 32.62(0.11) | 31.48(0.38) | 33.43(0.36) | 34.25(0.69) | 32.78(0.47) | 36.85(0.51) | |
| NTC | - | - | - | - | - | - | - | - | - | - | |
Average Ct values of sensitivity assays using 10-fold serial dilutions of a multi-target artificial positive control (APC) from 1 ng to 1 fg with primer and probe sets BruCo5F/5R/5P, BosCo8F/8R/8P, DecCo11F/11R/11P, ObsCo12F/12R/12P and PeaCo14F/14R/14P.
R2, linear correlation; Y, slope; Ex, reaction efficiency; SD, standard deviation; the Ct (threshold cycle) values are average of three replicates.