TABLE 1.

Sequence alignment and primer design for the detection of Aeromonas sp. virulence genes

Organism	Nucleotide sequence accession no.	% Similaritya	Primer combinationb	Site locations	% Similarity by PCR-ASAc	No. of PCR-RFLP fragmentsd
A. hydrophila	GBeM84709	100.0	C1, AHCF1-AHCR1	1661–1682, 1871–1892	100.0g	3
A. hydrophila	EMhX65045	96.9	C1, AHCF1-AHCR1	919–940, 1871–1892	100.0	3
A. hydrophila	GB M16495	86.9	C2, AHCF1-AHCR2	1309–1330, 1519–1540	92.2	3
A. hydrophila	EM X65043	84.7	C2, AHCF1-AHCR2	1203–1224, 1413–1433	90.5	3
A. hydrophila	EM X65044	84.7	C2, AHCF1-AHCR2	1202–1223, 1413–1433	90.1	3
A. sobria	EM X65046	74.4	C3, AHCF1-AHCR3	1117–1138, 1327–1348	76.7	2
A. salmonicida	EM X65048	74.7	C3, AHCF1-AHCR3	1119–1140, 1329–1350	76.7	2
A. caviae	GB U40711	77.5	C4, AHCF2-AHCR4	985–1006, 1195–1216	84.1	2
A. sobria	EM Y00559	73.6	C5, AHCF3-AHCR5	1639–1660, 1849–1870	81.5	4
A. trota	GB AF064068	73.1	C5, AHCF3-AHCR5	1174–1195, 1383–1405	81.9	4

^aCompared to the 2,528 bp of AHCYTOEN (GenBank nucleotide sequence accession no. M84709). 

^bPrimer sequences with mismatched bases underlined: AHCF1, 5′-GAGAAGGTGACCACCAAGAACA-3′; AHCR1, 5′-AACTGACATCGGCCTTGAACTC-3′; AHCF2, 5′-GAGAAGGTCTCCACCAAGAACA-3′; AHCR2, 5′ AGCTGACATCGGCCTTGAACTC-3′; AHCF3, 5′-GAGAAGGTCTCGACCAAGAACA-3′; AHCR3, 5′-AATTGACCTCGGCCTTGAACTC-3′; AHCR4, 5′ AGCTGACATCCGCCTTGAACTC-3′; AHCR5, 5′ AGCTCATATCCGCTTTGAACTC-3′. 

^cCompared to the 232-bp amplicon sequence of the AHCYTOEN gene. 

^dPCR-RFLP of 232 bp using HpaII. 

^eGB, GenBank. 

^fPrimer combination number. 

^gValue of 100.0% = 232-bp amplicon size. 

^hEM, European Molecular Biology Laboratory.