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Development of indirect competitive enzyme-linked immunosorbent and immunochromatographic strip assays for carbofuran detection in fruits and vegetables

Yao, Leijun, Liu, Liqiang, Song, Shanshan, Kuang, Hua, Xu, Chuanlai
Food and agricultural immunology 2017 v.28 no.4 pp. 639-651
apples, carbofuran, cell fusion, cross reaction, cucumbers, enzyme-linked immunosorbent assay, fruits, haptens, immunization, immunoaffinity chromatography, mice, monoclonal antibodies, rapid methods, toxicity
Carbofuran is a highly toxic pesticide used in fruits and vegetables. In this study, we produced a specific and sensitive monoclonal antibody (mAb) which was prepared based on a hapten that was derivatized with benzofuranol against carbofuran. Following mice immunization and cell fusion, we obtained three monoclonal cell lines: 6D5, 3H2, and 3C6. The cell line 3H2 generated mAb with the highest affinity and sensitivity. The half maximal inhibitory concentration was 0.3 ng/mL, and the cross-reactivity was <1%. Based on this mAb, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and immunochromatographic test strip (ICS) assay were developed to detect carbofuran residues in cucumbers and apples. The working range of ic-ELISA was 0.1–1 ng/mL, and the cutoff value of ICS was 1 ng/mL. The analytical recovery of carbofuran in cucumber and apple samples ranged from 81% to 97%. Both methods represent rapid screening tools for carbofuran detection in fruits and vegetables.