Table 2.
Performance of the Phylloxera LAMP assay using the optimised primer ratio and concentration (1:6:3; 0.4 µM). (a) Phylloxera specimens from different life stages, “crude” DNA (non-destructive) and “clean” DNA (destructive) extractions. (b) Phylloxera “crude” DNA (non-destructive) extracts from all life stages, blind panel test (workshop results).
| Specimens† | n | Amplification time (min) | Anneal derivative temperature (oC) | ||||
|---|---|---|---|---|---|---|---|
| Mean | SD | Min–Max | Mean | SD | Min–Max | ||
| a | |||||||
| Adult | 6 | 11.0 | 1.0 | 10.0–12.8 | 79.7 | 0.2 | 79.5–79.9 |
| Crawler | 6 | 10.5 | 0.6 | 10.0–11.5 | 79.8 | 0.2 | 79.6–80.0 |
| Egg | 6 | 10.3 | 1.0 | 9.0–11.5 | 79.7 | 0.1 | 79.6–79.7 |
| G1–G38* | 7 | 11.3 | 0.9 | 10.5–11.5 | 79.7 | 0.2 | 79.5–79.9 |
| Positive (G4) | 3 | 11.8 | 0.5 | 11.3–12.3 | 79.4 | 0.3 | 79.1–79.7 |
| b | |||||||
| Egg | 6 | 12.4 | 1.0 | 11.0–14.0 | 79.5 | 0.3 | 79.1–79.8 |
| Crawler | 3 | 13.8 | 0.4 | 13.5–14.3 | 79.5 | 0.3 | 79.3–79.8 |
| Intermediate | 5 | 13.5 | 2.0 | 12.3–17.0 | 79.4 | 0.2 | 79.2–79.6 |
| Adult | 10 | 12.1 | 0.6 | 11.0–13.0 | 79.3 | 0.2 | 79.1–79.6 |
| Alate | 1 | 14.3 | 0.0 | 14.3 | 79.4 | 0.0 | 79.4 |
| Positive (G 30) | 5 | 11.5 | 0.9 | 10.0–12.3 | 79.5 | 0.5 | 79.1–80.3 |
†Adult, crawler, egg, intermediate, adult and alate “crude” DNA extract (G4 strain) HotSHOT HS6 method. G1–G38, *seven strains G1, G4, G7, G19, G20, G30 and G38 of adult phylloxera “clean” DNA extract, Qiagen kit.