分析化学2024，Vol.52Issue(6)：828-837,中插20-中插23,14.DOI:10.19756/j.issn.0253-3820.231112
溴氰菊酯酶联免疫吸附分析和胶体金免疫层析分析方法的建立
Enzyme Linked Immunosorbent Assay and Gold Nanoparticle-based Lateral Flow Immunoassay for Rapid Detection of Deltamethrin
摘要
Abstract
In this study,nine hybridoma cells secreting monoclonal antibodies against deltamethrin were prepared,and the monoclonal antibody 4D4E11 with best sensitivity was selected to develop indirect enzyme-linked immunosorbent assay (ic-ELISA) and gold nanoparticle-based lateral flow immunoassay (LFIA) for detection of deltamethrin. The optimal working buffer for ic-ELISA was 0.01 mol/L phosphate buffer (pH 7.4) containing 0.2 mol/L NaCl and 20％ methanol,while 0.01 mol/L phosphate buffer (pH 7.4) containing 1 mol/L NaCl,5‰Tween-20 and 10％methanol for LFIA. Under the optimal conditions,the half inhibition concentration (IC50) and limit of detection (IC10) of ic-ELISA were 10.60 ng/mL and 1.43 ng/mL respectively,and the limit of detection of the developed LFIA was 0.5μg/mL. The developed ic-ELISA and LFIA showed no cross-reactivities (CRs) with eight kinds of analogues of deltamethrin,which indicated the excellent specificity of proposed immunoassays. The average recoveries of the ic-ELISA in spiked tomato,cabbage and lettuce samples were 79.8％-92.6％with relative standard deviations of 0.8％-5.5％. The detection results of LFIA were consistent with the spiked concentrations in the range of 1-5 mg/kg. Meanwhile,the results of ic-ELISA and LFIA showed close correlation with high performance liquid chromatography (HPLC) in the test of blind lettuce samples. The experimental results demonstrated that the two immunoassays proposed here were suitable for rapid detection of deltamethrin with high sensitivity and high accuracy.关键词
溴氰菊酯/单克隆抗体/间接竞争酶联免疫吸附法/胶体金免疫层析法Key words
Deltamethrin/Monoclonal antibody/Indirect competitive enzyme-linked immunosorbent assay/Gold nanoparticle-based lateral flow immunoassay引用本文复制引用
崔盼盼,王贵绣,吴艳玲,张召贤,丁园,华修德..溴氰菊酯酶联免疫吸附分析和胶体金免疫层析分析方法的建立[J].分析化学,2024,52(6):828-837,中插20-中插23,14.基金项目
国家自然科学基金项目(No.32102258)和安徽省重点研究与开发计划项目(No.2022i01020009)资助.Supported by the National Natural Science Foundation of China(No.32102258)and the Anhui Provincial Key Research and Development Program(No.2022i01020009). （No.32102258）
