Development of real-time PCR method for rapid detection and quantification of Heterosigma akashiwo

To rapidly detect the harmful algae H. akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H. akashiwo were used for designing species-specific primers, and a RFQ-PCR （Realtime Fluorescent Quantitative Polymerase Chain Reaction） method was developed for quantitative detection of H. akashiwo. Primer H. akashiwo and TaqMan probe were designed, and the specificity of primer was checked with PCR. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other H. akashiwo samples, which were assayed with both the RFQ- PCR method and visual count under microscope.