發(fā)布時間：2018-03-14 10:39

  本文選題：東海原甲藻　切入點：中肋骨條藻　出處：《中國海洋大學》2014年碩士論文　論文類型：學位論文

【摘要】：近年來，，赤潮的頻繁出現(xiàn)對經(jīng)濟和海洋生態(tài)系統(tǒng)造成了巨大影響，破壞了海洋生態(tài)系統(tǒng)平衡和穩(wěn)定。解析赤潮暴發(fā)的機制，構(gòu)建檢測、預(yù)警和防止赤潮暴發(fā)的方法和措施成為海洋生態(tài)研究的重點。應(yīng)用ELISA檢測方法，利用特定酶與特異性抗體連接的特性，通過測定酶促反應(yīng)底物的吸光度值可以實現(xiàn)對目的抗原的測定。ELISA檢測技術(shù)具有較高的靈敏度和重現(xiàn)性，被廣泛的應(yīng)用于微生物檢測。在赤潮藻鑒定領(lǐng)域也有所應(yīng)用。 本研究主要針對近年在東海海域引起赤潮暴發(fā)現(xiàn)象的東海原甲藻（Prorocentrum donghaiense）和中肋骨條藻（Skeletonema costatum）作為研究對象，制備針對這兩種藻細胞蛋白的高特異性的單克隆抗體和多克隆抗體，建立快速、準確、穩(wěn)定的分析和鑒定赤潮藻的免疫學方法。 本研究采用細胞免疫及雜交瘤細胞技術(shù)成功的制備了針對東海原甲藻和中肋骨條藻的單克隆抗體和多克隆抗體，建立了針對這兩種藻的雙抗體ELISA定量檢測方法；最低檢出限度為1×103cells/mL。用雙抗體夾心ELISA定量檢測方法與顯微鏡計數(shù)法對單一藻種以及模擬混合藻種進行檢測實驗，結(jié)果發(fā)現(xiàn)兩者的檢測結(jié)果高度一致。最后，用本研究建立的抗體定量檢測方法對東海赤潮監(jiān)控海區(qū)和嵊泗海水浴場的海水樣品進行現(xiàn)場檢驗，結(jié)果表明，本研究建立的針對東海原甲藻和中肋骨條藻的雙抗體夾心ELISA檢測方法，能夠快速完成對這兩種赤潮藻的定量檢測。 本研究建立的雙抗體夾心ELISA檢測方法可以快速、穩(wěn)定、準確的特異性檢測和識別海水中的東海原甲藻和中肋骨條藻，不會受到其他海洋浮游藻類以及較小懸浮顆粒物的干擾。應(yīng)用該方法對海水中的東海原甲藻和中肋骨條藻進行檢測，可以實現(xiàn)對赤潮暴發(fā)的快速檢測預(yù)報和防治控制。
[Abstract]:In recent years, the frequent occurrence of red tide has a great impact on the economy and marine ecosystem, and has destroyed the balance and stability of marine ecosystem. Methods and measures for early warning and prevention of red tide outbreaks have become the focus of marine ecological research. Using ELISA detection method, using specific enzyme linked to specific antibodies, By measuring the absorbance value of the substrate of enzymatic reaction, the detection of target antigen. Elisa has high sensitivity and reproducibility, and has been widely used in microbial detection, as well as in the field of identification of red tide algae. This study was aimed at Prorocentrum donghaiensee and Skeletonema costatum, which have caused red tide outbreaks in the East China Sea in recent years, and prepared highly specific monoclonal antibodies and polyclonal antibodies against these two kinds of algae cell proteins. To establish a rapid, accurate and stable immunological method for the analysis and identification of red tide algae. In this study, monoclonal antibodies and polyclonal antibodies against Prorocentrum chinensis and P. costatum were successfully prepared by cell immunity and hybridoma cell techniques, and a double antibody ELISA quantitative detection method was established for the detection of these two species of algae. The lowest detection limit is 1 脳 103 cells / mL. The double antibody sandwich ELISA quantitative detection method and microscope counting method are used to detect the single algae species and the simulated mixed algae species. The results show that the detection results are highly consistent. The antibody quantitative detection method established in this study was used to detect the sea water samples in the red tide monitoring area of the East China Sea and Shengsi bathing site. The results showed that, In this study, a double antibody sandwich ELISA method was developed for the detection of Prorocentrum and Ribella costatum in the East China Sea, which can rapidly complete the quantitative detection of these two species of red tide algae. The double antibody sandwich ELISA assay developed in this study can be used to detect and identify Prorocentrum East China Sea and S. costatum in seawater quickly, stably and accurately. This method can be used to detect the Prorocentrum of the East China Sea and the Ribella costatum in the sea water, which can be used to detect and forecast the red tide outbreak quickly and to control the red tide outbreak.
【學位授予單位】：中國海洋大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：X55;X835

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