Table 3.
Validation and Comparison of the VCN Determination by Different Strategies ACM and SCG-DP-PCR
| Validation 1 | Validation 2 | Validation 3 | |
|---|---|---|---|
| Ct (DP-CAR) | 24.77 ± 0.03 | 24.28 ± 0.03 | 25.31 ± 0.02 |
| Ct (DP-RNaseP) | 24.93 ± 0.05 | 24.80 ± 0.01 | 25.50 ± 0.02 |
| ΔCt | −0.16 | −0.52 | −0.19 |
| VCN/cell (SCG-DP-PCR) | 2.2 | 2.9 | 2.3 |
| VCN/cell (ACM) | 1.3 | 2.3 | 1.5 |
| Copies/Std. (ACM) | Ct (Validation 1) | Ct (Validation 2) | Ct (Validation 3) |
| 3 × 105 | 20.9 ± 0.1 | 21.4 ± 0.1 | 21.5 ± 0.1 |
| 3 × 104 | 24.2 ± 0.1 | 24.2 ± 0.1 | 24.9 ± 0.2 |
| 3 × 103 | 27.5 ± 0.1 | 27.6 ± 0.1 | 27.6 ± 0.0 |
| 3 × 102 | 30.9 ± 0.0 | 31.1 ± 0.1 | 31.6 ± 0.3 |
| 30 | 33.1 ± 0.1 | 34.0 ± 0.3 | 35.3 ± 0.6 |
Three independent experiments were performed (validation 1, 2, and 3). Mean Ct values from qPCR were used for linear regression (ACM) and ΔCt calculation (SCG-DP-PCR). Reactions were performed in triplicates. Ct values are represented as mean ± SD.