微囊藻毒素时间分辨荧光免疫分析方法

目的建立高灵敏的微囊藻毒素(MC-LR)的时间分辨荧光间接竞争免疫分析方法(MC-LR-TRFIA)。方法以MC-LR与牛血清白蛋白的偶联物(MC-LR-BSA)包被96孔板为固相抗原,与游离MC-LR共同竞争有限的抗MC-LR多克隆抗体;用稀土离子Eu3+标记的羊抗兔抗体进行示踪。结果该方法的灵敏度为0.01μg/L,测量范围为0.01~20μg/L,ED80、ED50和ED20分别为0.16、0.57和1.70μg/L。平均回收率为101%,与MC-LF和MC-RR平均交叉反应分别为0.73%和35%。比较MC-LR-TRFIA和MC-LR-ELISA检测MC-LR,两者的相关系数为0.958。研究表明,MC-LR-TRFIA是目前报导的MC-LR检测中最灵敏的方法。结论该方法适用于水样中MC-LR的定量检测。

The use of ultrasensitive time-resolved fluoroimmunoassay for rapid detection of microcystin LR

OBJECTIVE: In order to provide a rapid and selectivity method for the determination of microcystin LR (MCLR). METHODS: An indirect competitive time-resolved fluoroimmunoassay (TRFIA) was developed. MC-LR-BSA was coated by physical adsorption onto the microtitre plate, MC-LR or sample with MC-LR as a competitor. Both them were incubated with limited anti- MC-LR antibody. and a goat antirabbit IgG-Eu3+ conjugate was used as a tracer. RESULTS: The sensitivity of MC-LR-TRFIA was 0.01 microg/L, and the recovery rate was 99.7%. RSD of CBL-TRFIA were 3.9%. The sensitivity of MC-LR -TRFIA provided a linear reaponse from 0.01 - 20 microg/L, with ED50 of 0.57 microg/L or ED80 of 0.16 microg/L and ED20 of 1.70 microg/L. The cross reactivity of the MC-LR-TRFIA with MC-LF and MC-RR was 0.73% and 35%. Both MC-LR-TRFIA and MC-LR-ELISA test were applied for the quantitative measurement of MC-LR in the same samples, and the coefficient of correlation was 0.958. CONCLUSION: The method is suitable to determine MC-LR in water samples.