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为建立快速、准确的赤潮藻鉴定和定量检测方法,以红色裸甲藻(Gymnodinium sanguineum)作为研究对象,将快速检测手段--实时荧光定量PCR(RFQ-PCR)技术应用于赤潮藻的种类鉴定中.首先以红色裸甲藻18S rDNA序列为靶区域,设计出适合用于RFQ-PCR技术的红色裸甲藻引物和TaqMan探针,并通过引物PCR验证确定其种特异性;进而应用实时荧光定量PCR方法建立红色裸甲藻快速检测方法.与显微镜计数方法比较,两者所获结果无显著性差异,且该方法的检测限远低于红色裸甲藻爆发赤潮时的藻密度,因此该方法可有效地应用于红色裸甲藻的检测,从而为红色裸甲藻赤潮的预警预测奠定基础.
In order to establish a rapid and accurate method for the identification and quantitative detection of red tide algae, Gymnodinium sanguineum was used as the research object, and the rapid detection method-real-time fluorescence quantitative PCR (RFQ-PCR) was applied to the species identification of red tide algae In this paper, we designed the primers for the detection of the genomic DNA of Gymnothochia durianus 18S rDNA with the primers RFQ-PCR and the real-time fluorescence Quantitative PCR method was used to establish the rapid detection method of Gymnodinium dabryanus.With the method of microscope counting, there was no significant difference between the two methods, and the detection limit of this method is far lower than that of algae The method can be effectively applied to the detection of the red dinoflagellate, thus laying a foundation for the early warning and forecasting of the red tide.