The purified isolates were grown in the brain-heart infusion
broth overnight at 37 C and centrifuged at 10,000 g for 10 min. The
resulting pellets were washed twice with the sterile physiological
saline, resuspended in the distilled water, and then boiled for
10 min. The bacterial lysatewas used immediately for PCR or stored
at 20 C. The presence of the V. vulnificus hemolysin A (vvhA) gene
was determined using PCR method as described previously . Each primer pair produced a 519-bp amplicon from
the positive strain. PCR products were electrophoresised on a 1.8%
agarose gel at 120 V, stained with ethidium bromide, and
photodocumented.