微囊藻毒素-LR的生物素-亲和素-时间分辨荧光免疫分析法  

Determination of microcystin-LR by using biotin-avidin-time-resolved fluoroimmunoassay

作　　者：张艺[1] 黄飚[1] 金坚[2] 陈蕴[2] 赵春城[2] 张珏[1] 

机构地区：[1]江苏省原子医学研究所卫生部核医学重点实验室江苏省分子核医学实验室,无锡江苏214063 [2]江南大学药学院

出　　处：《环境与健康杂志》2015年第1期58-60,共3页Journal of Environment and Health

基　　金：江苏省社会发展基金(BE2009631)

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。Objective To develop an uhrasensitive 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, 1：1000 diluted anti-MC-LR polyclonal antibody,5 p^g/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 p^g/ml in water samples, while the RSDs of inner- and 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.

关 键 词：微囊藻毒素-LR 时间分辨荧光免疫分析法 生物素 亲和素 

分 类 号：O657.6[理学—分析化学]