| Microcystis is one genus of cyanobactreria which could cause water bloom ineutrophication water and most of them could produce microcystins. Microcystis has drownmore and more attention in recent years and seen as a big threat to aquatic ecosystem andwater supply security. The conventional analysis methods of microcystins, which includingbiologic analysis, physicochemical analysis and biochemical analysis, couldn not been usedto determine the potential microcystin-producing Microcystis. PCR-based methods couldeffectively detect the potential microcystin-producing Microcystis., while it could possiblegenerate false positive. Therefore, the development of effective systems for early monitoring,analysis and warning producing-microcystins Microcystis is of great significance.The development of modern biotechnology and the mass bioinformation made theestablishing of gene analysis methods possible. In this study, we established enzyme-linkedsandwich hybridization assay (ELSHA) which could directly detect the microcystinsynthetase gene The main contents and conclusions are summarized as follows:(1) The sequence of mcyJ gene of Microcystis was analyzed and the phylogenetic treeof mcyJ was constructed. The results showed that mcyJ gene is highly conservative andthus could be used for establishing the molecular biological detection method ofmicrocystin-producing Microcystis.(2) The capture probe and signal probe were designed and synthesized based on thesequence of mcyJ gene of Microcystis, and tested using ELSHA. It was found that theprobes are highly specific for detecting microcystin-producing Microcystis.(3) The ELSHA was established based on the sequence of mcyJ gene of Microcystisand optimized the experimental hybridization conditions. The reliability and accuracy ofELSHA was proved by detecting several microcystis cultured in lab.(4) The concentration of microcystin-producing Microcystis in water samples fromYujia Lake and South Lake were detected by ELSHA, and the results were discussed andcompared with other test indexes of water samples. |
