Table 2.
Primers and probe sequences used to detect brood diseases and parasites.
| No. | Target | Primer | Sequence (5’→3′) | Target gene | Reference |
|---|---|---|---|---|---|
| 1 | Paenibacillus larvae (AFB) | AFB-F | AAATCATCATGCCCCTTATG | 16S rRNA, 158 bp | This study |
| AFB-R | CGATTACTAGCAATTCCGACT | ||||
| Probe | FAM-CGTACTACAATGGCCGGTACAACG–BHQ-1 | ||||
| 2 | Melissococcus plutonius (EFB) | EFB-F | TGTTGTTAGAGAAGAATAGGGGAA | 16S rRNA, 69 bp | This study |
| EFB-R | CGTGGCTTTCTGGTTAGA | ||||
| Probe | Cy5-AGAGTAACTGTTTTCCTCGTGACGGT-BHQ-3 | ||||
| 3 | Apocephalus borealis (phorid fly) | Phorid-F | CCTCTGTTCTACTTTCATTGGTTTAT | 18S rRNA, 255 bp | This study |
| Phorid-R | GAGRGCCATAAAAGTAGCTACACC | ||||
| Probe | JOE-GGCATTAGTATTACGACGCGAGAGGTG–BHQ-1 | ||||
| 4 | Acarapis woodi mite | ACAR-F | CAGTAGGGCTAGATATCGATACCCGAGCTT | COI, 247 bp | This study |
| ACAR-R | TGAGCTACAACATAATATCTGTCATGAAGA | ||||
| Probe | TexasRed-CAATCCACCTACAGAAAATAAAAATAAAAATCC–BHQ-2 | ||||
| 5 | Nosema cerana | Nosema-F | CGGATAAAAGAGTCCGTTACC | LSU rRNA, 249 bp | This study |
| Nosema-R | GAGCAGGGTTCTAGGGAT | ||||
| Probe | FAM-CGTTACCCTTCGGGGAATCTTC–BHQ-1 | ||||
| 6 | Aspergillus flavus (stonebrood) | ASP-F | GCTGCCCATCAAGCACGG | ITS2, 127 bp | This study |
| ASP-R | CCTACAGAGCGGGTGACAAAG | ||||
| Probe | JOE-TGTGTGTTGGGTCGTCGTCCCCTCTC–BHQ-1 | ||||
| 7 | Ascosphaera apis (chalkbrood) | ASCO-F | ATTGCGCCCTCTGGTATTC | ITS2, 215 bp | This study |
| ASCO-R | CCACTAGAAGTAAATGATGGTTAGA | ||||
| Probe | TexasRed-GCTTGAGGGTTGCAATGACGCTCG BHQ-2 |
Abbreviations: AFB: American foulbrood; EFB: European foulbrood.