Table 2.
Primer sequences and PCR conditions for each virulence gene target.
| Gene | Strains ID | Primer sequence (5′-3′) | Hybridization temperature (°C) | Elongation time (min) |
|---|---|---|---|---|
| actA | All strains | Forwarda, GTATTAGCGTATCACGAGGA | 60 | 1 |
| Reversea, CAAGCACATACCTAGAACCA | ||||
| Forwardb, AAGMGTCAGTTRYGGATRCT | 57 | 1 | ||
| Reverseb, CCCGCATTTCTTGAGTGTTT | ||||
| hly | 227, 728, 872, 873, 874, 875, 876 | Forwarda, GGCCCCCTCCTTTGATTAGT | 60 | 2 |
| Reversea, GCCTCTTTCTACATTCTTCACAAA | ||||
| 355 | Forwarda, TATGCTTTTCCGCCTAATGG | 57 | 1 | |
| Reversea, CGTGTGTGTTAAGCGGTTT | ||||
| 250, 338, 877, 878 | Forwarda, AAAAGAGAGGGGTGGCAAAC | 60 | 2 | |
| Reversea, GCCTCTTTCTACATTCTTCACAAA | ||||
| All strains | Forwardb, CCAGGTGCTCTCGTRAAAGC | 57 | 1 | |
| Reverseb, RCCGTCGATGATTTGAACTT | ||||
| mpl | All strains | Forwarda, GCCACCTATAGTTTCTACTGCAAA | 57 | 1 |
| Reversea, TGRAGAATTAAKTTTTCTCTAACATTT | ||||
| All strains | Forwardb, ATACGCTCGCGCTAAGTTCT | 60 | 1 | |
| Reverseb, GCTTCTTATTCGCCCATCTCG | ||||
| plcA | All strains | Forwarda, ATCAAAGGAGGGGGCCATT | 60 | 1 |
| Reversea, CCGAGGTTGCTCGGAGATATAC | ||||
| plcB | All strains | Forwarda, ATTGGCGTGTTCTCTTTAGGc | 57 | 1 |
| Reversea, CAAAGAAAAAGATTAACCTCCCTTT | ||||
| prfA | All strains | Forwarda, TTCAGGTCCKGCTATGAAAC | 57 | 1 |
| Reversea, AACTCCATCGCTCTTCCAGA |
a
External primers located in upstream and downstream regions surrounding the targeted gene.
b
Internal primers.
c
Roche et al., 2005.