东海原甲藻抗体的制备及酶联免疫检测方法的建立

Preparation of antibodies and development of an enzymelinked immunosorbent assay for detection of Prorocentrum donghaiense

通过制备针对东海原甲藻细胞破碎物的多克隆和单克隆抗体,建立了基于双抗体酶联免疫分析定量检测东海原甲藻(Prorocentrum donghaiense)的检测方法。利用该方法对单一藻种、混合藻种和现场样品中的东海原甲藻进行检测的结果与镜检结果相一致,最低检测限度为1×103 cells/m L。该方法的建立对中国近海赤潮暴发的实时监控具有重要意义。

Antibody-based methods are developed to detect planktonic microalgae both qualitatively and quantitatively recent years. Herein, we prepared both polyclonal and monoclonal antibodies against Prorocentrum donghaiense and then set up the method of double-antibody ELISA to quantitatively detect P. donghaiense. The specificity and validity of the double-antibody ELISA technique were evaluated with cultured pure strains, mixed strains and field samples, and by comparison with microscopy observations. The results showed that there was no significant difference between the two methods, and the minimum detection limit was 1×103 cells/m L. The double-antibody ELISA technique provides a convenient tool for rapid assessment of HAB species in marine environments, which has great significance for performance of a real-time monitoring on the outbreaks of harmful algal bloom in China coastal sea.