Development of an Heterologous Immunoassay for Ciprofloxacin Residue in Milk

被引:9
作者
Jiang Jinqing [1 ]
Zhang Haitang [1 ]
An Zhixing [1 ]
Xu Zhiyong [1 ]
Yang Xuefeng [1 ]
Huang Huaguo [1 ]
Wang Ziliang [1 ]
机构
[1] Henan Inst Sci & Technol, Coll Anim Sci, Xinxiang 453003, Henan, Peoples R China
来源
INTERNATIONAL CONFERENCE ON SOLID STATE DEVICES AND MATERIALS SCIENCE | 2012年 / 25卷
关键词
Ciprofloxacin; Artificial antigen; polyclonal antibody; Indirect competitive ELISA; Heterologous; Milk; LINKED-IMMUNOSORBENT-ASSAY; INDIRECT COMPETITIVE ELISA; ENROFLOXACIN; FLUOROQUINOLONES; ANTIBIOTICS; ANTIBODIES; PRODUCTS;
D O I
10.1016/j.phpro.2012.03.318
中图分类号
T [工业技术];
学科分类号
08 ;
摘要
A heterologous immunoassay has been developed for the determination of Ciprofloxacin (CPFX) residues in milk. For this reason, carbodiimide active ester method was employed to synthesize the artificial antigen of CPFX-BSA, and mixed anhydride reaction was used to prepare the coating antigen of CPFX-OVA to pursue the heterologous sensitivity. Based on the square matrix titration, an icELISA method was developed for the quantitative detection of CPFX in cattle milk. The dynamic range was from 0.036 to 92.5 ng/mL, with LOD and IC50 value of 0.019 ng/mL and 1.8 ng/mL, respectively. Except for a high cross-reactivity (89.7%) to Enrofloxacin, negligible cross-reactivity to the other compounds was observed. After optimization, 0.03 mol/L of HCl, or 10% of methanol was used in the assay buffer. 20-fold dilution in cattle milk gave an inhibition curve almost the same as that in PBS buffer. The regression equation for this assay was y = 0.9036 x + 1.4574, with a correlation coefficient (R-2) of 0.9844. The results suggest the veracity of the heterologous immunoassay for detecting CPFX residue in milk.
引用
收藏
页码:1829 / 1836
页数:8
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