时间分辨荧光免疫一步法检测微囊藻毒素的研究

One Step Time-Resolved Fluorescence Immunoassay for Direct Estimation of Microcystins

利用稀土元素Eu^3＋标记微囊藻毒素偶联物MCLR—BSA，在固相包被二抗的微孔板上建立了微囊藻毒素的直接竞争时间分辨荧光免疫法．条件优化后Eu^3＋标记物的稀释度为1：50，微囊藻毒素单抗的合适浓度为100ng／mL．该法对微囊藻毒素检测的灵敏度为0．02ng／mL，试剂的测量范围是0．05～10ng／mL，回收率达到94％以上．

A direct-competitive time-resolved fluorescence immunoassay（TRFIA） for microcystin detection was established, which was based on europium labeled MCLR-BSA conjugant and microtiter plates coated with anti-mouse IgG. The optimal dilution of europium labeled MCLR-BSA conjugant is 1/50 and most appropriate titration of anti-microcystin-LR （MCLR） monoclonal antibodies is 100 ng/mL. The standard curve under the optimal conditions shows that the quantitative range is from approximately 0.05 ng/mL to 10 ng/mL The detection limit of the method is 0.02 ng/mL for MCLR, recoveries for microcvstin （ 〉 94% ） in auantitative range is satisfactorv.