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期 刊 ：植物保护学报 2008年

关键词：鱼藤酮；半抗原；抗体；酶联免疫吸附测定；

Keywords:rotenone, hapten, antibodies, ELISA,

摘 要 ：为建立鱼藤酮免疫分析方法,以鱼藤酮为原料合成4-{2-肼基}-4-丁酸(简称鱼藤酮腙琥珀酸半酯,RH-HS)半抗原,通过碳化二亚胺法(EDC)、活酯法(AE)将半抗原与牛血清蛋白(BSA)偶联制备免疫原Ⅰ、Ⅱ。将免疫原分别免疫BALB/c鼠获得相应的抗体Ⅰ和抗体Ⅱ,测定抗体冻干粉效价,分别为4.10×10⁵和8.29×10⁵。并进行抗原抗体的竞争抑制能力、特异性和鱼藤酮的添加回收试验。结果表明:抗体Ⅰ、Ⅱ对鱼藤酮最低检测限(IC₂₀)分别为0.440μg/mL和0.437μg/mL。两抗体对吡蚜酮、克百威的交叉反应率均小于1.1%,抗体Ⅰ对4个与鱼藤酮结构相类似化合物的交叉反应率均高于56%,较抗体Ⅱ高。鱼藤酮的免疫分析技术(ELISA)与HPLC检测手段相比,在检测线性范围和灵敏度方面,两者差异不明显;在样品前处理方面,ELISA显示出快速、简单、有效的优点。

Abstract:In order to establish the immunoassay method of rotenone, the hapten 4-{2-hydrazinyl}-4-oxobutanoic acid(RH-HS) was synthesized from rotenone. The hapten was conjugated to bovine serum albumin (BSA), with the 1-ethyl-3-3-dimethylaminopropyl carbodiimide (EDC) method and active ester (AE) method to prepare immuneogenⅠand immuneogenⅡ. Corres- ponding antibodies were prepared through the immuneogens in BALB/c mouse. Titers of antibody Ⅰand antibody Ⅱ were 4.10×10⁵ and 8.29×10⁵, respectively. With the experiments about the affinity, specificity of antibodies and the recoveries of rotenone, the results showed that the limit of detection (IC₂₀) for rotenone were 0.440μg/mL and 0.437μg/mL. The percentages of cross-activity to pymetrozine and carbofuran were less than 1.1%, and for the other 4 similar compounds, the antibody Ⅰwas much higher than antibody Ⅱ with the percentage of 56% above. The ELISA which was established by the antibodyⅠ, was content with detecting the similar compounds of rotenone. There was not diversity to compare with HPLC in lineal range and sensitivity. But they had displayed a rapid, simple, effective method in sample preparation.

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