烯酰吗啉人工抗原的合成及其ic-ELISA方法的建立  

Synthesis of Dimethomorph Artificial Antigen and Establishment of Indirect Competitive-Enzyme Linked Immunosorbent Assay

作　　者：冯荣虎[1] 吴会玲 王珍妮[1] 张恒 张世伟[1] FENG Ronghu;WU Huiling;WANG Zhenni;ZHANG Heng;ZHANG Shiwei(Shenzhen Academy of Metrology&Quality Inspection,Shenzhen 518060,China)

机构地区：[1]广东省深圳市计量质量检测研究院,广东深圳518060

出　　处：《食品科技》2022年第9期264-270,共7页Food Science and Technology

基　　金：深圳市基础研究专项(自然科学基金)基础研究面上项目(YCYJ20170817162416978)。

摘　　要：目的:旨在建立一种可定量检测果蔬中烯酰吗啉残留的间接竞争酶联免疫吸附检测(Indirect competitive-enzyme linked immunosorbent assay,ic-ELISA)方法。方法:首先合成烯酰吗啉半抗原,通过活泼酯法与钥孔血蓝蛋白(Keyhole limpet hemocyanin,KLH)及牛血清白蛋白(Bovine albumin,BSA)偶联获得免疫抗原DMM-KLH和包被抗原DMM-BSA;通过杂交瘤技术获得1株能够稳定分泌烯酰吗啉的单克隆抗体细胞株DMM-2C4,并通过腹水制备大量抗体。建立烯酰吗啉的ic-ELISA检测方法。结果:烯酰吗啉IC_(50)为2.05 ng/mL,检测限为0.35 ng/mL,与其他真菌杀菌剂无交叉反应。在果蔬中的添加回收率为68.3%~115.5%。结论:该方法能有效应用于果蔬中烯酰吗啉的快速检测。Objective:This study aimed to establish an indirect competitive enzyme-linked immunosorbent assay for quantitative detection of Dimethomorph residues in fruits and vegetables.Method:The prepared dimethomorph haptens were conjugated to keyhole limpet hemocyanin as immungens(DMM-KLH)and bovine serum albumin as coating antigens(DMM-BSA)via active ester method.A stable hybridoma cell line DMM-2C4 was obtained by immunizing Balb/c mice.Large amounts of monoclonal antibodies were prepared by inducing the ascites of mice.An indirect competitive-enzyme linked immunosorbent assay(ic-ELISA)was developed.Result:The results showed that the IC_(50) of ic-ELISA was 2.05 ng/mL,the detection limit was 0.35 ng/mL.It had no cross reaction with other fungicide.The recoveries of dimethomorph from spiked fruits and vegetables by ic-ELISA were from 68.3%to 115.5%.Conculsion:This result suggested that the developed ic-ELISA has good sensitivity and accuracy for the detection of dimethomorph residues in vegetable samples.

关 键 词：烯酰吗啉 间接竞争酶联免疫吸附法(ic-ELISA) 人工抗原 

分 类 号：TS207.3[轻工技术与工程—食品科学]