Abstract
[Objective] To establish a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for detecting enrofloxacin (ENR) residues in animal-derived food. [Methods] ENR was conjugated with keyhole limpet hemocyanin (KLH) using the carbodiimide method to prepare an immunogen, and New Zealand white rabbits were immunized to produce high-sensitivity antibodies. Based on this, the detection system was optimized to establish ic-ELISA, the detection performance was evaluated, and spiked recovery tests were conducted in animal-derived food. [Results] Under the optimized conditions of coating antigen concentration at 0.25 μg/mL, antibody dilution of 1∶40 000, secondary antibody dilution of 1∶5 000, primary antibody incubation for 15 min, secondary antibody incubation for 30 min, and color development for 5 min, the IC50 of the method was 27.76 pg/mL. The detection range was 0.49~1 577.29 pg/mL, and the limit of detection was 0.33 pg/mL. The intra- and inter-assay coefficients of variation were both less than 15%. The method showed good specificity in trace detection of ENR, with recovery rates of 93.37%~97.06% in chicken and 87. 15%~97.59% in fish. [Conclusion] This method demonstrates excellent sensitivity, precision, specificity, and accuracy, and provides an effective tool for detecting trace residues of ENR in animal-derived food. 康怀彬 （1963 —），男，河南科技大学教授，硕士。E-mail: khbin001@163.com
Publication Date
5-13-2026
First Page
37
Last Page
44
DOI
10.13652/j.spjx.1003.5788.2025.80103
Recommended Citation
Jiaxiang, HUANG; Yanfei, LI; Kaile, WANG; Min, ZHANG; Yao, WANG; and Huaibin, KANG
(2026)
"Indirect competitive enzyme-linked immunosorbent assay for the detection of enrofloxacin residues in animal-derived food,"
Food and Machinery: Vol. 42:
Iss.
3, Article 5.
DOI: 10.13652/j.spjx.1003.5788.2025.80103
Available at:
https://www.ifoodmm.cn/journal/vol42/iss3/5
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