Table 2.
Primer sequences and PCR amplification conditions for the detection of Vibrio virulent genes.
| Target gene | Primer sequence (5ʹ-3ʹ) | Amplicon size (bp) | PCR cycling conditions | Reference |
|---|---|---|---|---|
| tdh | F: GTAAAGGTCTCTGACTTTTGGAC R: TGGAATAGAACCTTCATCTTCACC |
270 | Initial denaturation at 94°C for 3 minutes, 30 cycles of denaturation at 94°C for 1 minute, primer annealing at 58°C for 1 minute, elongation at 72°C for 1 minute, and a final strand elongation at 72°C for 5 minutes. | [10] |
| trh | F: TTGGCTTCGATATTTTCAGTATCT R: CATAACAAACATATGCCCATTTCCG |
486 | ||
|
| ||||
| ctxAB | F: GCCGGGTTGTGGGAATGCTCCAAG R: GCCATACTAATTGCGGCAATCGCATG |
536 | Initial denaturation at 94°C for 10 minutes, 30 cycles of denaturation at 94°C for 1 minute, primer annealing at 59°C for 1 minute, elongation at 72°C for 2 minutes, and a final strand elongation at 72°C for 10 minutes. | [41] |
| zot | F: TCGCTTAACGATGGCGCGTTTT R: AACCCCGTTTCACTTCTACCCA |
947 | ||
|
| ||||
| flrA | F: GAGGCAACAGCACCATCAAA R: CGCATCTATATCAGGGACAA |
503 | Initial denaturation at 95°C for 5 minutes, 35 cycles of denaturation at 95°C for 45 seconds, primer annealing at 72°C for 45 seconds, and a final strand elongation at 72°C for 15 minutes. | [8] |
| vpsR | F: GGTGAGTAGCCATAAGCAAG R: CATCCAGCACCACAGTATCT |
911 | ||