Study Of Molecular Biology Detection Technique Of Prorocentrum Lima

Prorocentrum lima is a common benthic dinoflagellate, also camp planktonic life, is the main source of diarrhetic shellfish poisoning(DSP), can cause human food poisoning by food chain, has already attracted much attention. In order to reduce the harm of P. lima, first, we must fully grasp whether P. lima exist in the natural water and its quantity, so study of a rapid detection technique of P. lima is very important.This study choose the area have significant difference from other phytoplankton of 18S rDNA area of P. lima, designed the primer fit for fluorescence quantitative PCR of P. lima, and validated the specificity of primer, application of SYBR Green real-time fluorescence quantitative PCR technique established the method to qualitative and quantitative detect P. lima rapidly, draw the quantitative counting standard curve, and validated the accuracy of the method with samples known cell quantity.The results show that, in theory, P. lima can be detected by this method when the sample contains four cells, has a high sensitivity; in the study of practical ecological investigation, measured value is 505(±14.72)cells/L(microscopic value is 600cells/L), error is 15.83%, meet the plankton investigation require error less than 20% in the ocean survey specification, Fully meet the require of the forecasting of toxic algae in the ecology study.This method has a high specificity, accuracy and sensitivity, the operation is simple, time-saving, can complete in 3h, remain highly relevant for the large samples detection, can serve as a method for P. lima qualitative and quantitative detection in the marine environmental monitor.