Table 5. Reproducibility of simultaneous detection of ApMV, ASPV and ASGV with multiplex RT-qPCR.
| Samples | Multiplex real-time PCR mean Ct values | |||
|---|---|---|---|---|
| ApMV | ASPV | ASGV | ||
| Three biological replicates of triple infected B71 RNA dilutions | B71 | 25.07 (0.29) | 25.30 (0.66) | 25.23 (0.72) |
| 10−1 | 28.82 (0.53) | 27.77 (0.15) | 29.10 (0.19) | |
| 10−2 | 31.05 (0.26) | 30.78 (0.09) | 32.84 (0.36) | |
| 10−3 | 34.26 (0.36) | 33.20 (0.38) | 37.17 (0.31) | |
| 10−4 | 37.07 (0.12) | 36.62 (0.28) | Na | |
| Two individual runs of serial dilutions of RNA transcripts | 106 copies | 25.03 (0.4) | 24.07 (0.12) | 31.4 (0.71) |
| 105 copies | 28.40 (0.49) | 27.41 (0.34) | 34.34 (0.19) | |
| 104 copies | 30.74 (0.35) | 29.81 (0.73) | 35.58 (0.06) | |
| 103 copies | 32.58 (0.49) | 32.04 (0.74) | 37.23 (0.49) | |
| 102 copies | 36.67 (0.85) | 35.88 (0.92) | Na | |
Reproducibility of the method as evaluated through calculation of mean Ct values and standard deviations (in brackets) of serial dilutions of the triple infected B71 RNA in three biological replicates and of RNA transcript serial dilutions in two individual runs. Serial 10-fold dilutions were prepared into RNA extracted from virus-negative tested pear tissue.
Na: not applicable