摘要
以化合物2-甲基-4-三氟甲基-5-噻唑甲酸(MTCA)为噻呋酰胺的半抗原合成人工抗原,采用活泼酯法合成免疫原MTCA-BSA,分别采用活泼酯法、混合酸酐法和N,N'-羰基二咪唑/4-二甲氨基吡啶(CDI/DMAP)法合成3种包被原MTCA-OVA-1、MTCA-OVA-2和MTCA-OVA-3。免疫动物后,最终以包被原MTCAOVA-3筛选并获得了具有高特异性的噻呋酰胺多克隆抗体,建立了检测噻呋酰胺的间接竞争酶联免疫吸附(ic-ELISA)分析方法。本方法线性检测范围为0.08~10.00 mg/L,半数抑制浓度(IC_(50))为1.39 mg/L,检出限为0.08 mg/L,对自来水、湖水和小麦中的噻呋酰胺添加回收率在72.0%~128.3%之间,检测结果与HPLC法具有良好的相关性(R^2=0.9994)。本研究建立的ic-ELISA方法可用于环境水样与小麦等农产品中噻呋酰胺残留的快速检测。
In the present study,2-methyl-4-(trifluoromethyl)thiazole-5-carboxylic acid(MTCA)was acted as hapten of thifluzamide to synthesize artificial antigens.Immunogen MTCA-bovine serum albumin(BSA)was synthesized by active ester method.Meanwhile,three coating antigens,MTCA-OVA-1,MTCA-OVA-2and MTCA-OVA-3were synthesized by active ester method,mixed anhydride method and N,N-carbonyldiimidazole/4-dimethylaminopyridine(CDI/DMAP)method,respectively.Anti-thifluzamide polyclonal antibody with high specificity was obtained from the immunized mice,then indirect competitive enzyme linked immunosorbent assay(ic-ELISA)method for thifluzamide detection was developed by using MTCA-OVA-3and polyclonal antibody.The linear detection range in ic-ELISA was0.08-10mg/L with an IC50of1.39mg/L and a LOD(IC10)of0.08mg/L.The recoveries for spiked samples including tap water,lake water and wheat ranged from72.0%-128.3%in ic-ELISA method.Good correlation(R2=0.9994)was obtained between the results of ic-ELISA and those of HPLC analysis.The proposed ic-ELSA was promising for rapid detection of thifluzamide in water and agricultural products.
作者
李潇潇
丁书茂
杨旭
袁均林
LI Xiao-Xiao;DING Shu-Mao;YANG Xu;YUAN Jun-Lin(Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science,Central China Normal University, Wuhan 430079, China)
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2018年第1期27-32,共6页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金项目(No.21577045)资助~~
关键词
噻呋酰胺
人工抗原
多克隆抗体
酶联免疫吸附分析
Thifluzamide
Artificial antigen
Polyclonal antibody
Enzyme linked immunosorbent assay
