Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction.

ABSTRACT

Vibrio anguillarum, an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum. PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum. Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA, or from six colony-forming units (CFU) mL(-1) of cultured V. anguillarum. However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g(-1) tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.