DefinePK

Abstract

Background:   In a previous report, we showed that  Vibrio shilonii  was found on whiteleg shrimp ( Litopenaeus   vannamei ) with acute hepatopancreatic necrosis disease in Thua Thien Hue province, Vietnam. This study was performed to develop a diagnostic molecular marker generated by random amplified polymorphic DNA (RAPD) for  V. shilonii  rapid detection. Methods:  Pathogen  Vibrio  spp. were isolated from shrimps and fishes, and were identified by 16S rRNA sequencing. Genetic diversity of  Vibrio  strains was analysis by RAPD technique. Specific PCR product for  V. shilonii  was cloned and sequenced. SCAR marker was developed from specific PCR product. Result:  Twenty random primers were evaluated for RAPD to identify DNA polymorphisms between  Vibrio  species. The random primer OPN-06 generated a 468-bp DNA fragment specific for  V. shilonii . This was then converted into a sequence-characterized amplified region (SCAR) marker designated N6-441. Conclusion:  Specific primers (Vshi-441F/Vshi-441R) amplified a unique DNA fragment in all  V. shilonii  isolates but not in the other  Vibrio  spp. This PCR assay showed significantly sensitive to the target DNA and reliably for the amplification the  V. shilonii  genome. Keywords: AHPND; RAPD; SCAR;  Vibrio shilonii ; Vietnam