溴氰菊酯残留酶联免疫分析方法的建立

在前期设计合成了溴氰菊酯半抗原（DM）和人工抗原（DM-BSA和DM-OVA）的基础上,进一步利用人工抗原（DM-BSA）免疫新西兰大白兔获得了溴氰菊酯多克隆抗体。研究表明,新西兰大白兔产生的抗体对溴氰菊酯产生了灵敏的特异性免疫反应,所得多克隆抗体的效价为25600。以DM-OVA为包被原建立溴氰菊酯间接竞争ELISA检测方法,确定了抗原抗体最适工作浓度均为1∶12800。在含10%甲醇、pH7.4、盐浓度0.1mol·L^-1的缓冲液条件下制作了溴氰菊酯的标准抑制曲线,抑制中浓度IC50为3.999μg·mL^-1,检出限IC10为0.023μg·mL^-1,检测范围为0.015-100μg·mL^-1。抗体对包括氯菊酯、氯氰菊酯、甲氰菊酯在内的8种菊酯类农药的交叉反应率较低,在苹果中的添加回收率为86.2%-105.8%。成功制备出溴氰菊酯特异性抗体,并建立了水果中溴氰菊酯残留间接竞争酶联免疫分析方法。

Development of Enzyme Immunoassays for Deltamethrin

On the base of preliminary studies, including the synthesis of hapten （DM） and the preparation of ariificial antigen （DM-BSA, DM-OVA）, polyclonal antibody against deltamethrin was successfully obtained by immunization of New Zealand white rabbits with DM- BSA. The highest titer of the antiserum was 25 600. As coating antigen, DM-OVA was used to establish an indirect competitive enzyme linked immunosorbent assay （IC-ELISA） for deltamethrin. The dihttion of the coating antigen and the antibody used in the ELISA were 1：12 800. Furthermore, the buffer solution used in ELISA with 10 % methanol, pH 7.4, 0.1 mol·L^-1 NaCl were also determined. The results indicated that the antibody produced from New Zealand white rabbits immunized had high affinity to deltamethrin tested by IC-ELISA, the IC50 for dehamethrin was 3.999μg·mL^-1 and the lowest detection limit ICl0 was 0.023 μg·mL^-1, the working range was assigned to concentrations 0.015-100 ）μg·mL^-1 for deltamethrin. Different cross-reactions with the closely related to deltamethrin, including cypermethrin, perme- thrin, lambda-cyhalothrin, fenpropathrin, bio-allethrin, tetramethrin, fenvalerate, empenthrin, were observed. There were low cross reactivities. Recovery rate of dehamethrin spiked to apple was 86.2%-105.8%. Special antibody was produced successfully and IC-ELISA was developed for the analysis of deltamethrin residue in fruits.