Table 1.
Oligonucleotide primers, PCR protocols, and amplicon sizes for each cPCR assay.
| Bacteria, amplified gene | Primer sequences (5′-3′)a | Cycling conditions | Amplicon size (bp) | Primer reference |
|---|---|---|---|---|
| Mannheimia haemolytica, lkt | F: GTCCCTGTGTTTTCATTATAAG | 95°C, 5 min; (94°C, 30 s; 58°C, 45 s; 72°C, 60 s) ×35 cycles; 72°C, 10 min | 385 | (22) |
| R: CACTCGATAATTATTCTAAATTAG | ||||
| Pasteurella multocida, 23S rRNA | F: GGCTGGGAAGCCAAATCAAAG | 95°C, 5 min; (94°C, 30 s; 58°C, 45 s; 72°C, 60 s) ×35 cycles; 72°C, 10 min | 1,432 | (23) |
| R: CGAGGGACTACAATTACTGTAA | ||||
| Histophilus somni, 16S rRNA | F: GAAGGCGATTAGTTTAAGAG | 95°C, 5 min; (94°C, 30 s; 55°C, 45 s; 72°C, 60 s) ×35 cycles; 72°C, 10 min | 400 | (24) |
| R: TTCGGGCACCAAGTRTTCA | ||||
| Mycoplasma bovis, uvrC | F: TTACGCAAGAGAATGCTTCA | 95°C, 5 min; (94°C, 30 s; 56°C, 45s; 72°C, 60 s) ×35 cycles, 72°C, 10 min95°C, 5 min; (94°C, 30 s; 56°C, 45s; 72°C, 60 s) ×35 cycles, 72°C, 10 min | 171 | (25) |
| R: TCATCCAAAAGCAAAATGTTAAA | ||||
| Mycoplasma bovis, 16S | F: GGGAGCAAACAGGATTAGATACCCT | 269 | ||
| R: TGCACCATCTGTCACTCTGTTAACCT |
a
F, forward primer; R, reverse primer.