【摘要】 目的运用生物素-亲和素信号放大系统,建立高灵敏的微囊藻毒素-LR(MC-LR)时间分辨荧光免疫分析(TRFIA)检测方法。方法采用5μg/ml二抗Ig G包被板,样品中的MC-LR与生物素-MC-LR半抗原(1∶300)竞争限量抗MC-LR抗体(1∶1 000),用铕标记的链霉亲和素示踪,采用TRFIA法检测水样中的MC-LR含量。结果在0.1~5 ng/ml的线性范围内,得到MC-LR-TRFIA的回归方程为y=-29 132x+38 057,r=0.991 5。方法的灵敏度为0.02 ng/ml,平均回收率为80.2%~102.0%,批内和批间的RSD分别为4.3%~8.1%和9.9%~15.2%。该方法检测MC-LR与MC-YR和MC-LF的交叉率均小于10%。结论基于生物素-亲和素信号放大系统的MC-LR-TRFIA方法灵敏度高、特异性好、操作简便,适于水中MC-LR浓度的检测。更多还原

【Abstract】 Objective To develop an ultrasensitive time- resolved fluoroimmunoassay( TRFIA) for the determination of microcystin-LR(MC-LR) with biotin-avidin amplified system. Methods 1∶300 diluted biotin-MC-LR-BSA,1∶1 000 diluted anti-MC-LR polyclonal antibody,5 μg/ml coated goat-anti-rabbit antibody and europium-labeled streptavidin were used in competitive TRFIA method, and MC-LR was detected in the water samples. Results The regression equation of MC-LR-TRFIA was y=-29 132x+38 057,r=0.991 5, in the linear range of 0.1-5 ng/ml. This method offered a sensitivity of 0.02 ng/ml.The average recovery rates were 80.2%-102.0% at spiked levels of 0.1-1 μg/ml in water samples, while the RSDs of innerand inter-assay were 4.3%-8.1% and 9.9%-15.2%,respectively. The cross-reactivity rates of MC-YR and MC-LF to MC-LR were both below 10%. Conclusion The method developed in the study is of high sensitivity,specificity,easy operation and applicable to the screening of MC-LR in samples.更多还原