Quantitative detection of Vibrio alginolyticus strain XSBZ14 by a newly developed RT-PCR method

Abstract Purpose Coral degradation is a worldwide ecological problem. Bacterial diseases are great danger to coral health. The pathogenic bacterium Vibrio alginolyticus XSBZ14 isolated from diseased had been identified as the of Porites andrewsi White syndrome (PAWS) in Xisha Archipelago on transmission experiment. To date, molecular mechanism by which this pathogen causes disease unknown, and diagnostics for caused have not developed. In an effort restore damaged ecosystems South China Sea, efforts underway transplant flat-branch shore corals. There therefore urgent need further develop specific rapid detection methods V. order prevent epidemic ensure successful implementation compilation transplants. Methods At first, low sequence identity single-copy S2 was selected genome in-house Perl script. Using designed primers, four different types standard curves were subsequently plotted accurate quantification strain samples (DNA, bacterial suspension, tissue, seawater). Then, use infect Galaxea fascicularis test culture water during week. Results method bacteria RT-PCR established. limit (LOD) 0.88 pg/reaction (0.44 pg/μL) DNA, 2 CFU/reaction (1000 CFU/mL) 20 seawater XSBZ14, respectively. addition, according results RT-PCR, could survive week, also be detected its reared seawater. Conclusion These indicated that robust tool pathogen, very useful PAWS epidemiological survey pathogen-free transplantation Sea. And other species their habitats might act important reservoir mediated horizontal reefs.