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Detection of Heterotrophic Dinoflagellate Pfiesteria piscicida (Dinophyceae) in Surface Water Samples Using Real-time PCR

  • Park, Tae-Gyu (Marine Ecology Research Division, National Fisheries Research and Development Institute) ;
  • Kang, Yang-Soon (Marine Ecology Research Division, National Fisheries Research and Development Institute) ;
  • Seo, Mi-Kyung (Marine Ecology Research Division, National Fisheries Research and Development Institute) ;
  • Park, Young-Tae (Marine Ecology Research Division, National Fisheries Research and Development Institute)
  • Published : 2008.12.31

Abstract

Heterotrophic dinoflagellate Pfiesteria piscicida (Dinophyceae) has been claimed to produce potent ichthyotoxins that cause disorientation and eventually death of fish and other marine animals. A real-time PCR probe targeting for SSU rRNA gene was used for detection of P. piscicida in Chinhae Bay, Korea. PCR inhibitors were successfully removed by dilution of template DNA. Positive detections were shown from surface water samples indicating the presence of P. piscicida in Chinhae Bay.

Keywords

References

  1. Bowers, H.A., T. Tengs, H.B. Glasgow, J.M. Burkholder, P.A. Rublee and D.W. Oldach. 2000. Development of real-time PCR assays for rapid detection of Pfiesteria piscicida and related dinoflagellates. Appl. Environ. Microbiol., 66, 4641-4648 https://doi.org/10.1128/AEM.66.11.4641-4648.2000
  2. Gibson, U.E.M., C.A. Heid and P.M. Williams. 1996. A novel method for real-time quantitative RT-PCR. Genome Res., 6, 995-1001 https://doi.org/10.1101/gr.6.10.995
  3. Heid, C.A., J. Stevens, K.J. Livak and P.M. Williams. 1996. Real-time quantitative PCR. Genome Res., 6, 986-994 https://doi.org/10.1101/gr.6.10.986
  4. Holland, P.M., R.D. Abramson, R. Watson and D.H. Gelfand. 1991. Detection of specific polymerase chain reaction product by utilizing the 5'-3' exonu-clease activity of Thermus aquatics DNA polymerase. Proc. Natl. Acad. Sci., USA, 88, 7276-7280
  5. Mumford, R.A., K. Walsh, I. Barker and N. Boonham. 2000. Detection of potato mop top virus and tobacco rattle virus using a multiplex real-time fluorescent reverse-transcription polymerase chain reaction assay. Phytopathology, 90, 448-453 https://doi.org/10.1094/PHYTO.2000.90.5.448
  6. Park, T.G., E.M. Bell, P. Imojen, P.A. Rublee, C.J.S. Bolch and G.M. Hallegraeff. 2007a. Detection of a novel ecotype of Pfiesteria piscicida (Dinophyceae) in an Antarctic saline lake by real-time PCR. Polar Biol., 30, 843-848 https://doi.org/10.1007/s00300-006-0244-0
  7. Park, T.G., M.F. de Salas, C.J.S. Bolch and G.M. Halle-graeff. 2007b. Development of a real-time PCR probe for quantification of the heterotrophic dinoflagellate Cryptoperidiniopsis brodyi (Dinophyceae) in environ-mental samples. Appl. Environ. Microbiol., 73, 2552-2560 https://doi.org/10.1128/AEM.02389-06
  8. Rublee, P.A., D.L. Reminfton, E.F. Schaefer and M.M. Marshall. 2005. Detection of the dinozoans Pfiesteria piscicida and P. shumwayae: A review of detection methods and geographic distribution. J. Eukaryot. Microbiol., 52, 83-89 https://doi.org/10.1111/j.1550-7408.2005.05202007.x
  9. Saito, K., T. Drgon, J.A.F. Robledo, D.N. Krupatkina and G.R. Vasta. 2002. Characterization of the rRNA Locus of Pfiesteria piscicida and development of standard and quantitative PCR-based detection assays targeted to the nontranscribed spacer. Appl. Environ. Micro-biol., 68, 5394-5407 https://doi.org/10.1128/AEM.68.11.5394-5407.2002
  10. Schena, L., F. Nigro and A. Ippolito. 2002. Identification and detection of Rosellina necatrix by conventional and real-time Scorpion-PCR. Eur. J. Plant Pathol., 108, 355-366 https://doi.org/10.1023/A:1015697813352
  11. Schena, L. and A. Ippolito. 2003. Rapid and sensitive detection of Rosellinia necatrix in roots and soils by real time Scorpion-PCR. J. Plant Pathol., 85, 15-25