Table 1 Primers and probe for qPCR and ddPCR used in this study
PCR assay | Institution/ country | Detection method | Target | Primer/probe position | Sequence (5’-3’) | Amplicon size (bp) | Reference |
|---|---|---|---|---|---|---|---|
qPCR1 | AUCVM/US | FRET qPCR | pol | Fw, 2321–2345 Rv, 2484–2507 Anchor probe, 2407–2430 Reporter probe, 2431–2454 | CCTCAATTCCCTTTAAACTAGAACG ATGGGCTTTGTAAGAGCATTTGTA GACGGGCCAGGCAATAATCCAGT-6-FAM LCRed640-TTCCCGGTACGGAAACCAAATGG-phosphate | 187 | |
qPCR2 | CentralStar/US | Multiplex TaqMan qPCR | pol, β-actin, spike-in control | N/A | N/A | N/A | [25] |
qPCR3 | LFDA-MG/ Brasil | TaqMan qPCR | LTR | BLV.LTR.114.Fw, 37–54 BLV.LTR.114.Rv, 131–150 BLV.LTR.114.Probe, 108–130 | GCCCCGTAAACCAGACAG CTCAGGGTGTGGATTTCTCG FAM-TACCTCCCC/ZEN/AACTTCCCCTTTCC-IABkF | 114 | [26] |
qPCR4 | CIVETAN/Argentina | SYBRGreen qPCR | pol | Fw, 4618–4636 Rv, 4657–4676 | CACCATTCACCCCCACTTG TCAGAGCCCTTGGGTGTTTC | 59 | [27] |
qPCR5 | FA-IU/Japan | Duplex TaqMan qPCR | pol, RPPH1 | N/A | N/A | N/A | [28] |
ddPCR6 | UdelaR/Urugway | TaqMan ddPCR | env | Fw, 5384–5406 Rw, 5525–5546 Probe, 5477–5497 | CAGTGACTGGGTTCCCTCTGTC AGGGCGAGRCCGGGTCCAGAG HEX-CCCTCCCTGGGCTCCCGARA-BHQ1 | 162 | [29] |
qPCR7 | VEINST/Croatia | TaqMan qPCR | pol | MRBLVL Fw, 2321–2340 MRBLVR Rv, 2421–2440 MRBLV Probe, 2341–2360 | CCTCAATTCCCTTTAAACTA GTACCGGGAAGACTGGATTA 6FAM-GAACGCCTCCAGGCCCTTCA-BHQ1 | 120 | [30] |
qPCR8 | INTA/Argentina | SYBRGreen qPCR | pol | BLVpol5f Fw, 2321–2340 BLVpol3r Rv, 2421–2440 | CCTCAATTCCCTTTAAACTA GTACCGGGAAGACTGGATTA | 120 | |
qPCR9 | LCV/Spain | Duplex TaqMan qPCR | pol β-actin | MRBLVL Fw, 2321–2340 MRBLVR Rv, 2421–2440 Probe, 2341–2360 ACT-1005-F Fw, 1005–1029 ACT-1135-R Rv, 1135–1114 ACT-1081 Probe, 1081–1105 | CCTCAATTCCCTTTAAACTA GTACCGGGAAGACTGGATTA Cy5-GAACGCCTCCAGGCCCTTCA-BHQ1 CAGCACAATGAAGATCAAGATCATC CGGACTCATCGTACTCCTGCTT HEX-TCGCTGTCCACCTTCCAGCAGATGT-BHQ1 | 120 130 | |
qPCR10 | NVRI/Poland | TaqMan qPCR | pol | MRBLVL Fw, 2321–2340 MRBLVR Rv, 2421–2440 MRBLV Probe, 2341–2360 | CCTCAATTCCCTTTAAACTA GTACCGGGAAGACTGGATTA 6FAM-GAACGCCTCCAGGCCCTTCA-BHQ1 | 120 | [30] |
qPCR11 | Anses/France | TaqMan qPCR | pol | MRBLVL Fw, 2321–2340 MRBLVR Rv, 2421–2440 MRBLV Probe, 2341–2360 | CCTCAATTCCCTTTAAACTA GTACCGGGAAGACTGGATTA 6FAM-GAACGCCTCCAGGCCCTTCA-BHQ1 | 120 | [30] |