Development of Rapid Analysis and Monitoring system for Microcystin

采用蛋白磷酸酶2A抑制法对环境水样中的微囊藻毒素进行快速分析。建立了一种从微囊藻细胞中快速提取微囊藻毒素的热处理方法，用蛋白磷酸酶2A抑制法对微囊藻毒素进行定量分析需要2小时。该方法准确、重现性好。将培养的微囊藻悬液进行蛋白磷酸酶2A抑制试验和商品化的酶联免疫吸附试验试剂盒检测，结果相似。但该方法的灵敏度不如酶联免疫吸附试验试剂盒。从微囊藻细胞中提取微囊藻毒素时，加热温度为50~60℃，加热时间大于20min。用-淀粉酶、纤维素酶和溶菌酶进行热处理对微囊藻毒素的提取有一定的促进作用；采用蛋白磷酸酶2A抑制法结合热处理法对天然水中的微囊藻毒素进行检测。浓缩部分对提取的微囊藻毒素进行超滤浓缩。检测部分采用蛋白磷酸酶固定化抑制法进行检测。在本研究中，饮用水源水和自来水样品中未检测到任何类型的微囊藻毒素。农业池塘样品中检测到高浓度的微囊藻毒素。微囊藻毒素的浓度与微囊藻细胞数和/或水温无关。

The fast analysis of microcystin in environmental water was examined by using a protein phosphatase 2A inhibition assay. The heat treatment method was developed for rapid extraction of microcystin from Microcystis cells.Quantitative analysis of microcystin by the protein phosphatase 2A inhibition assay required two hours. The assay was accurate and reproducible. The cultured Microcystis suspension was submitted to the protein phosphatase 2A inhibition assay as well as commercial ELISA assay kit with similar results. But sensitivity of this assay was not so good as the ELISA assay kit. The microcystins were extracted from Microcystis cells by heat conditions such as temperature of 50-60℃ and heating time of more than 20 minutes. The heat treatment with-amylase, cellulase and lysozyme was slightly facilitated to extraction of it.Uses of the protein phosphatase 2A inhibition assay with the heat treatment method facilitate investigation of microcystin in natural water.The Monitoring system for Microcystin was made in three parts, such as extraction, concentration and detection equipment.The microcystins were extracted from Microcystis cells by heat treatment in extraction part. Extracted microcystins were concentrated by ultra-filtration in concentration part. And, those were determined by inhibition assay use of immobilization of protein phosphatase in detection part.In this study, any type microcystins was not detected in samples of resource water for drinking and samples of tap water. And, high concentration of microcystins was detected in samples of pond for agriculture. Concentration of microcystins was not related with Microcystis cell Numbers and/or water temperature.