建立水产品中己烯雌酚残留ELISA检测方法

›› 2010, Vol. 37 ›› Issue (10): 43-47.

建立水产品中己烯雌酚残留ELISA检测方法

赵新宇1，王权1，闫叶娜2

（1.中国农业科学院上海兽医研究所兽医公共卫生实验室，上海 200241；2.南京农业大学，南京 210095）

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-10-20 发布日期:2010-10-20
通讯作者: 王权

Establishment of Enzyme-linked Immunosorbent Assay for Diethylstilbestrol

ZHAO Xin-yu1，WANG Quan1，YAN Ye-na2

(1.Laboratory of Veterinary Public Health，Shanghai Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Shanghai 200241，China；2.Nanjing Agricultural University，Nanjing 210095，China)

Received:1900-01-01 Revised:1900-01-01 Online:2010-10-20 Published:2010-10-20
Contact: WANG Quan

摘要/Abstract

摘要： 本试验利用己烯雌酚(DES)单克隆抗体，通过优化反应条件，加标回收试验，确定ELISA检测方法相关参数，建立了己烯雌酚残留的酶联免疫吸附（ELISA）检测法。试验结果显示，包被抗原最佳浓度为2 μg/mL，DES单克隆抗体最佳稀释倍数为1∶3×104；标准曲线呈线性相关，相关系数R2=0.9956，IC50=1.103 ng/mL，最适检测范围为0.125～128 ng/mL，检测敏感度为0.077 ng/g；批内和批间变异分别为6.14%和7.48%；鳝鱼肌肉组织的加标回收率为77.6%~94.6%；特异性试验结果表明，该单抗与己烯雌酚单羧丙基醚交叉反应率为112.7%，与其它水产禁用药物交叉反应率＜0.1%。

关键词: 己烯雌酚; 药物残留; 酶联免疫吸附法

Abstract: The enzyme-linked immunosorbent assay (ELISA) for the detection of diethylstilbestrol (DES) was established, the most suitable working conditions of ELISA were optimized. The optimal concentration of the coating antigen (DES-OVA) and optimal dilution of monoclonal antibody against DES were 2 μg/mL and 1∶3×104，respectively. The linear regression equation of the standard curve was R2=0.9956. The range of linearity was from 0.125 to 128 ng/mL. The 50% inhibition value (IC50) was 1.103 ng/mL, sensitivity of the method was 0.077 ng/g，respectively. The intra-assay and inter-assay CVs were 6.14% and 7.48%. The recoveries of mud eel ranged from 77.6% to 94.6%. The very high specificity was evaluated by the significant cross-reactivity with analogous compounds and other aquatic antibiotics，the McAb against DES had 112.7% cross-reactivity with diethylstilbestrol-mono-carboxylic propyl ether，there was no cross-reactivity with other aquatic antibiotics(＜0.1).

Key words: diethylstilbestrol; drugs residue; ELISA

中图分类号:

S859.84

赵新宇;王权;闫叶娜. 建立水产品中己烯雌酚残留ELISA检测方法[J]. , 2010, 37(10): 43-47.

ZHAO Xin-yu;WANG Quan;YAN Ye-na. Establishment of Enzyme-linked Immunosorbent Assay for Diethylstilbestrol[J]. , 2010, 37(10): 43-47.

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