Table 1.
Oligonucleotide primers used for PCR amplification of Vibrio species-specific gene fragments.
| Target organism | Primer sequence (5ʹ-3ʹ) | Targeted gene | Amplicon size (bp) | PCR cycling conditions | Reference |
|---|---|---|---|---|---|
| All bacterial strains (universal 16S rRNA gene sequence) | 27F: AGAGTTTGATCATGGCTCAG 1492R: GGTACCTTGTTACGACTT |
16S rRNA | 1420 | Initial denaturation at 94°C for 3 minutes, 25 cycles of denaturation at 94°C for 1 minute, primer annealing at 55°C for 1 minute, elongation at 72°C for 2 minutes, and a final strand elongation step at 72°C for 10 minutes. | [37] |
|
| |||||
| V. cholerae | F: CACCAAGAAGGTGACTTTATTGTG R: GGTTTGTCGAATTAGCTTCACC |
ompW | 304 | 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. | [38] |
| V. cholerae serogroup O1 | F: TCTATGTGCTGCGATTGGTG R: CCCCGAAAACCTAATGTGAG |
rfbO1 | 638 | ||
|
| |||||
| V. cholerae | F: AAGACCTCAACTGGCGGTA R: GAAGTGTTAGTGATCGCCAGAGT |
sodB (1) | 248 | Initial denaturation at 95°C for 10 minutes, 35 cycles of denaturation at 92°C for 40 seconds, primer annealing at 57°C for 1 minute, elongation at 72°C for 1.5 minutes, and a final strand elongation at 72°C for 10 minutes. | [39] |
| V. parahaemolyticus | F: GCAGCTGATCAAAACGTTGAGT R: ATTATCGATCGTGCCACTCAC |
flaE | 897 | ||
| V. vulnificus | F: GTCTTAAAGCGGTTGCTGC R: CGCTTCAAGTGCTGGTAGAAG |
hsp60 | 410 | ||
| V. mimicus | F: CATTCGGTTCTTTCGCTGAT R:GAAGTGTTAGTGATTGCTAGAGAT |
sodB | 121 | ||
|
| |||||
| V. harveyi | F: CTTCACGCTTGATGGCTACTG R: GTCACCCAATGCTACGACCT |
vhh | 235 | Initial denaturation at 95°C for 10 minutes, 30 cycles of denaturation at 95°C for 1 minute, primer annealing at 50°C for 1 minute, elongation at 72°C for 1 minute, and a final strand elongation at 72°C for 10 minutes. | [40] |