Table 2.
Conditions of the reactions as described in the original studies and using the standard protocol.
| Gibb et al. [9] | Huang et al. [10] | Panning et al. [11] | Standard Protocol Mekambo EBOV RNA | Standard Protocol EBOV Synthetic RNA | RealStar® Filovirus Altona Diagnostics | |
|---|---|---|---|---|---|---|
| Reverse Transcription | 55 °C/45 min | 50 °C/30 min | 50 °C/30 min | 50 °C/15 min | 50 °C/15 min | 55 °C/20 min |
| Denaturation | 94 °C/1 min | 94 °C/5 min | 95 °C/1.5 min | 95 °C/2 min | 95 °C/2 min | 95 °C/2 min |
| Cycling | 40 | 45 | 45 | 40 | 40 | 45 |
| Denaturation | 94 °C/15 s | 94 °C/15 s | 95 °C/15 s | 95 °C/15 s | 95 °C/15 s | 95 °C/15 s |
| Amplification | 60 °C/30 s | 60 °C/1 min | 72 °C/20 s | 60 °C/1 min | 60 °C/1 min | 72 °C/15 s |
| RNA volume | 5 µL | 1–3 µL | 3 µL | 5 µL | 10 µL | 10 µL |
| Total volume | 50 µL | 23–25 µL | 25 µL | 25 µL | 30 µL | 30 µL |
| Primers concentration | 0.5 µM | 0.4 µM | 0.2 µM Forward 0.3 µM Reverse |
0.4 µM Gibb 0.4 µM Huang 0.2 µM Panning |
0.625 µM Gibb 0.5 µM Huang |
- |
| Probes concentration | 0.2 µM | 0.1 µM | 0.0667 µM | 0.16 µM Gibb and Huang | 0.25 µM Gibb | - |
| 0.2 µM each Panning | 0.125 µM Huang |