Table 1 Reaction temperature optimization for RT-RPA-VF assays targeting the CDV N gene
Temperature | No. positive/ no. replications | ||||||||
|---|---|---|---|---|---|---|---|---|---|
Synthesized RNA transcript dilution (5 × copies/μl) | |||||||||
108 | 107 | 106 | 105 | 104 | 103 | 102 | 101 | N | |
44 °C | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
42 °C | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
39 °C | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 1/3 | 0/3 | 0/3 | 0/3 |
37 °C | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 2/3 | 0/3 | 0/3 |
35 °C | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 |
33 °C | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 1/3 | 0/3 | 0/3 |