Rapid detection and Quantification of Fish Killing Dinoflagellate Cochlodinium polykrikoides (Dinophyceae) in Environmental Samples Using Real-time PCR

摘要

The mixotrophic dinoflagellate Cochlodinium polykrikoides was reported to be linked to major fish kills in Korea and Japan since the 1990s. Rapid and sensitive detection of microalgae has been problematic because morphological identification of dinoflagellates requires light microscopic and scanning electron microscopic observations that are time consuming and laborious compared to real-time PCR. To address this issue, a real-time PCR probe targeting the ITS2 rRNA gene was used for rapid detection and quantification of C. polykrikoides. PCR inhibitors in water column samples were removed by dilution of template DNA for elimination of false-negative reactions. A strong association between cell quantification using real-time PCR and microscopic countssuggests that the real-time PCR assay is an alternative method for cell estimation of C. polykrikoides in environment samples.