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Objective To prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin (IgY) to build an enzyme-linked immunosorbent assay (ELISA) for citrinin (CTN). Methods CTN was conjugated with bovine serum albumin (BSA),ovalbumin (OVA) with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet (UV) spectrometry and Infrared (IR) spectrometry.Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).ELISA with IgY was established.Cross-reactivity of lgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied. Results UV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA.SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD.The indirect competitive ELISA showed that the detection limit of CTN synthesized.The established ELISA can be used to determine CTN- contaminated samples.