Development of two enzyme-linked immunosorbent assay formats for thifluzamide residues’ analysis based on distinct polyclonal antibodies
Li, Xiaoxiao | Zhao, Yun | Ding, Shumao | You, Huihui | Feng, Weiyong | Yang, Xu | Yuan, Junlin
Haptens 2-Methyl-4-(trifluoromethyl)thiazole-5-carboxylic acid and 2,6-Dibromo-4-(trifluoromethoxy)aniline, the two moieties of thifluzamide, were conjugated with carrier proteins for the synthesis of artificial antigens. Two distinct anti-thifluzamide polyclonal antibodies (PAb-1 and PAb-2) were produced from the immunized female Balb/c mice. The indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) in two formats based on the PAbs was developed for thifluzamide analysis. The concentration of 50% inhibition (IC₅₀) of ELISA-1 was 1.39 mg L⁻¹ and its limit of detection (LOD) was 0.082 mg L⁻¹. Meanwhile, ELISA-2 had a similar IC₅₀ of 1.96 mg L⁻¹ and a LOD of 0.074 mg L⁻¹ as ELISA-1. Both the raised PAbs exhibited high specificity to thifluzamide. The recoveries for spiked samples including water and wheat ranged from 72.0% to 128.4%, and the accuracy of ELISA was confirmed by high-performance liquid chromatography. In summary, the ic-ELISA might be a promising tool for simple, sensitive and rapid detection of thifluzamide residues in real samples.
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