TABLE 1.
Specific oligonucleotide primers for PCR amplification of the genes for C. perfringens toxins α, β, β2, and ɛ and enterotoxin
| Toxin/gene | Primer | Oligonucleotide sequence | Fragment length (bp) | Annealing temp (°C) | Ramping time (s)a | Reference or source |
|---|---|---|---|---|---|---|
| α/cpa | CPALPHATOX1-L | 5′-AAG ATT TGT AAG GCG CTT-3′ | 1,167 | 46 | 60 | 3 |
| CPALPHATOX1-R | 5′-ATT TCC TGA AAT CCA CTC-3′ | |||||
| β-cpb | CPBETATOX1-L | 5′-AGG AGG TTT TTT TAT GAA G-3′ | 1,025 | 39 | 90 | 3 |
| CPBETATOX1-R | 5′-TCT AAA TAG CTG TTA CTT TGT G-3′ | |||||
| β2/cpb2 | P319BETA2 | 5′-GAA AGG TAA TGG AGA ATT ATC TTA ATG C-3′ | 573 | 48 | This work | |
| P320BETA2 | 5′-GCA GAA TCA GGA TTT TGA CCA TAT ACC-3′ | |||||
| ɛ/etx | CPETOXIN1-L | 5′-AAG TTT AGC AAT CGC ATC-3′ | 961 | 46 | 60 | 3 |
| CPETOXIN1-R | 5′-TAT TCC TGG TGC CTT AAT-3′ | |||||
| Enterotoxin/cpe | CPENT1-L | 5′-TAACAATTTAAATCCAATGG-3′ | 933 | 46 | 60 | 3 |
| CPENT1-R | 5′-ATTGAATAAGGGTAATTTCC-3′ |
a
Ramping time from the annealing temperature to an extension temperature of 72°C.