Table 1.
Sensitivity and specificity of rt-qPCR Assays using hydrolysis probes and SYBR Green to quantify laboratory prepared dilutions of Plasmodium falciparum
| Dilutions (replicates) | Hydrolysis probe | SYBR Green | |
|---|---|---|---|
| Crossing point (SD), Quant value (SD) | |||
| In Water | 6.4 × 105 (×3) | 24.6 (0.06), 670 000 (28 000) | 20.5 (0.10), 740 000 (48 000) |
| 6.4 × 104 (×3) | 28.2 (0.09), 60 000 (3 700) | 24.6 (0.68), 57 000 (27 000) | |
| 6.4 × 103 (×3) | 30.8 (0.50), 9 200 (1 600) | 27.6 (0.38), 8 100 (1 600) | |
| 6.4 × 102 (×10) | 34.6 (0.40), 550 (140) | 32.9 (0.57), 550 (200) | |
| 6.4 × 101 (×10) | 37.9 (0.55), 56 (20) | 36.1 (1.46), 90 (70) | |
| 6.4 (×10) | 37.9 (n/a), 52 (n/a) | 40.39 (n/a), 4 (n/a) | |
| In Negative Extract | 6.4 × 105 (×3) | 24.7 (0.02), 650 000 (8 600) | 23.0 (0.22), 710000 (100 000) |
| 6.4 × 104 (×3) | 28.1(0.20), 62 000 (8 400) | 26.6 (0.21), 61 000 (8 200) | |
| 6.4 × 103 (×3) | 30.7 (0.41), 6 800 (1 200) | 30.1 (0.44), 7,600 (2 600) | |
| 6.4 × 102 (×10) | 33.5 (0.24), 660 (120) | 34.7 (0.37), 410 (120) | |
| 6.4 × 101 (×10) | 37.2 (1.09), 60 (50) | 37.2a (1.22), 90 (90) | |
| 6.4 (×10) | 37.6 (n/a), 40 (n/a) | 39.7a (1.75), 25 (45) | |
a Indicates nonspecific melting peaks of 79°C obtained by melting curve analysis of replicates tested.
n/a = Not Applicable
SD = Standard Deviation