Table 6.
Primers and probes for multiplex real-time RT-PCRa
| Viruses | Primers/probes | Sequences (5'-3') |
|---|---|---|
| BPI3 i | BPI3euro Forward | GGTAGGAGCACCTCCACGATT |
| BPI3euro Reverse | GCTCCAAGGCATGCTGGATA | |
| BPI3euroMGB | VIC-AAGATCTTGTTCACACATTC-MGB-NFQ | |
| BPI3 ii | BPI3 Forward2 | TGATTGGATGTTCGGGAGTGA |
| BPI3Reverse2 | AGAATCCTTTCCTCAATCCTGATATACT | |
| BPI3Fam | FAM-TACAATCGAGGATCTTGTTCA-MGB-NFQ | |
| BRSV [14] | BRSVn Forward | GGTCAAACTAAATGACACTTTCAACAAG |
| BRSVn Reverse | AGCATACCACACAACTTATTGAGATG | |
| BRSVLNACyan500 | Cyan500-TAGTACAGGTGACAA+CA+T+TG-BBQ | |
| BoHV-1 [15] | gB Forward | TGTGGACCTAAACCTCACGGT |
| gB Reverse | GTAGTCGAGCAGACCCGTGTC | |
| gBCy5 | Cy5-AGGACCGCGAGTTCTTGCCGC-BHQ2 | |
| β-actin | Bac1F_uni | GACAGGATGCAGAARGAGATCAC |
| Bac2R_uni | TCCACATCTGCTGGAAGGTG | |
| β-actin FAM-MGB | FAM-TGAAGATCAAGATCATCG-MGB-NFQ | |
a All the primers and probes are obtained from Applied Biosystems (Warrington, UK) except BRSV LNAcyan500 probe (TIB Molbiol,Berlin, Germany) and gBCy5 probe (Eurofins MWG Operon, Germany).
Abbreviations: BHQ black hole quencher, MGB-NFQ minor groove binding-non fluorescent quencher, LNA locked nucleic acid, BBQ Blackberry quencher.