Establishment Of Elisa Method For Detection Of Two Kinds Of Red Tide Algae

In recent years, Red Tides caused a tremendous impact on the economy andmarine ecosystems which occurred frequently and damaged the balance and stabilityof the marine ecosystem. Resolution mechanisms and causes of red tide phenomenon,building red tide detection, early warning and prevention methods and measures ofred tide outbreaks have become an important part of the marine ecology. Detected byELISA method, using the specific properties of the enzyme can be attached to theantibody specificity of enzymatic reaction by measuring the absorbance of thesubstrate to achieve the purpose of indirectly detecting the antigen. Because ofits good reproducibility, high sensitivity of enzymes, the reaction can be greatlyamplified effect, which can make the detection result showed a high sensitivity, betterresponse experiments. ELISA technology has been widely used in cell counts andspecies identification of red tide algae.In this study, we focused on Prorocentrum donghaiense and Skeletonemacostatum which cause the outbreak of red tide in the East China Sea phenomenon inrecent years as a research object, and then we made highly specific monoclonal andpolyclonal antibodies which for the two kinds of Red Tide.In this study, we successfully prepared for both monoclonal and polyclonalantibodies of the cell protein of Prorocentrum donghaiense and Skeletonema costatumwith the use of the immune cells and hybridoma technology.And established thesandwich ELISA quantitative detection methods for these two algae. The correlationcoefficient of quantitative detection standard curve established has good correlation;The minimum detection limit is1×103cells/mL; Not only for the single algae speciesbut also for the mixed species of algae, the results of antibody we established arebasically consistent with the results of microscopic detection; On the basis of theabove work, we use the method of ELISA to detect Prorocentrum donghaiense andSkeletonema costatum which is from the East China Sea and bathing water of Shengsi.We find the method of ELISA we established can accurately completed thequantitative analysis of the two kinds of red tide.The sandwich ELISA detection method developed in this study can be fast,stable and specific accurate detection and identification of Prorocentrum donghaienseand Skeletonema costatum in the seawater, without interference of other marinephytoplankton and small suspended particles.Using this methods, we can forecast theoutbreaks of red tide algae’s trend, and realize the fast monitoring, forecasting andprevention control of the red tide outbreak.