Table 4. Evaluation of the multiplex RT-qPCR.
s/n | Sample | Host | Geographic region | Symptoms | Multiplex RT-qPCR | Conventional RT-PCR | ||||
---|---|---|---|---|---|---|---|---|---|---|
ASPV | ASGV | ApMV | ASPV | ASGV | ApMV | |||||
1 | 6006 | Pear | Peloponnese | Fire blight symptomatology | 34.99 | 35.02 | - | - | - | - |
2 | G02 | P.amygdaliformis | Central Greece | Symptomless | - | - | - | - | - | - |
3 | 3492 | Apple | Peloponnese | Graft necrosis | 20.03 | 21.74 | 25.58 | + | + | + |
4 | B65 | Pear | Peloponnese | Reduced leaf size, short internodes | 23.93 | - | 36.66 | + | - | + |
5 | 3096 | Pear | Crete | Absence of vegetative development | - | - | - | + | - | - |
6 | 5847 | Pear | Thessaly | Reduced leaf size, chlorosis, stunting | 33.96 | 35.35 | - | + | - | - |
7 | KY1 | Quince | Central Greece | Symptomless | - | - | - | - | - | - |
8 | 2814 | Apple | Macedonia | Chlorosis, leaf curl | 22.87 | 23.93 | 27.64 | + | + | + |
9 | MT | Apple | Peloponnese | Symptomless | 21.52 | 23.93 | 26.51 | + | + | + |
10 | Tragic | Pear | Central Greece | Stunting | 33.7 | - | - | + | - | - |
11 | M4 | Apple | Central Greece | Symptomless | 23.41 | 23.63 | 27.17 | + | + | + |
12 | EVA2 | Apple | Central Greece | Symptomless | 21.84 | - | - | + | - | - |
13 | 2970 | Pear | Thessaly | Necrosis of branches | 33.14 | 35.44 | - | + | - | - |
14 | ELEA2 | Apple | Central Greece | Symptomless | 23.05 | - | 36.04 | + | - | + |
15 | WIL | Pear | Central Greece | Symptomless | - | - | - | - | - | - |
16 | 4176 | Pear | Central Greece | Symptomless | 29.83 | - | - | + | - | - |
17 | EP913 | Pear | Epirus | Wilting | 32.35 | 32.65 | 29.38 | + | - | + |
18 | B39 | Apple | Thessaly | Cankered branches | 20.56 | 23.71 | 27.34 | + | + | + |
19 | 4215 | Pear | Macedonia | Necrotic spots, stunting | 30.85 | 33.55 | - | + | - | - |
20 | EVP2 | Pear | Central Greece | Symptomless | - | - | - | + | - | - |
21 | 3248 | Pear | Central Greece | Leaf spots | 36.63 | - | - | - | - | - |
22 | B47 | Pear | Macedonia | Cankered limbs, shoot necrosis | 36.06 | - | - | - | - | - |
23 | PEL5 | Apple | Thessaly | Reduced fruit size | 24.65 | 24.04 | 20.88 | + | + | + |
24 | KPY | Pear | Central Greece | Symptomless | - | - | - | - | - | - |
25 | 3363 | Pear | Thrace | Reduced leaf and branch size | 35.82 | 36.53 | - | - | - | - |
26 | 4608 | Pear | Thessaly | Reduced fruit size | - | - | - | - | - | - |
27 | APL5 | Apple | Macedonia | Symptomless | 34.67 | 36.02 | 35.81 | - | - | + |
28 | 15643 | Pear | Macedonia | Limbs with cankers | - | - | - | - | - | - |
29 | B70 | Apple | Macedonia | Mosaic | 25.89 | 24.05 | 24.33 | + | + | + |
30 | M3 | Apple | Epirus | Symptomless | - | - | - | - | - | - |
31 | M1 | Apple | Central Greece | Symptomless | 24.33 | 25.87 | - | + | + | - |
32 | M2 | Apple | Central Greece | Symptomless | 22.15 | 24.99 | - | + | + | - |
33 | A1 | Pear | Central Greece | Symptomless | - | - | - | - | - | - |
34 | A2 | Pear | Central Greece | Symptomless | 26.63 | 25.95 | - | + | + | - |
35 | A3 | Pear | Central Greece | Symptomless | - | - | - | - | - | - |
36 | A4 | Pear | Central Greece | Symptomless | 25.3 | 26.28 | - | + | + | - |
Totala | 25 | 18 | 11 | 22 | 11 | 11 |
Total RNAs isolated from thirty-six samples of pome trees derived from commercial orchards, were analysed by the developed multiplex RT-qPCR and conventional RT-PCR assays. Comparison between methods was performed on the same RNA. Samples detected positive are indicated by cycle thresholds (Ct) and (+), for multiplex RT-qPCR and conventional RT-PCR, respectively.
a Number of samples in which at least one virus was detected