A heterologous indirect competitive enzyme linked immuno sorbent assay (icELISA) has been developed for the determination of enrofloxacin (ENR) residues in poultry. For this purpose, carbodiimide active ester method was employed to synthesize the artificial antigen of ENR- bovine serum albumin (BSA) while mixed-anhydride technique was used to synthesize the coating antigen of ENR- ovalbumin (OVA), to pursue the heterologous sensitivity. By square matrix titration, an icELISA method was developed, and the linear range was from 0.02 to 86.3 ng/mL, with limit of detection (LOD) and IC50 value of 0.8 and 0.01 ng/mL, respectively. After optimization, 5% of NaOH was used in the assay buffer and this ELISA system can tolerate methanol not higher than 30%. The correlation coefficients (R2) between concentration spiked and concentration determined were 0.9975 in chicken muscle and 0.9959 in duck muscle, respectively. Therefore, this assay has the potential to be incorporated into a quantitative monitoring program for the rapid screening of ENR residue in poultry muscles.
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机译：已开发出一种异源间接竞争酶联免疫吸附测定法（icELISA），用于测定家禽中的恩诺沙星（ENR）残留。为此目的，采用碳二亚胺活性酯法合成ENR-牛血清白蛋白（BSA）的人工抗原，同时采用混合酸酐技术合成ENR-卵清蛋白（OVA）的包被抗原，以寻求异源敏感性。通过正方形基质滴定，开发了一种icELISA方法，线性范围为0.02至86.3 ng / mL，检测限（LOD）和IC50值分别为0.8和0.01 ng / mL。优化后，在测定缓冲液中使用5％的NaOH，该ELISA系统可耐受不超过30％的甲醇。加标浓度与测定浓度之间的相关系数（R2）在鸡肌肉中为0.9975，在鸭肌肉中为0.9959。因此，该测定方法有可能被纳入定量监测程序中，以快速筛选家禽肌肉中的ENR残留物。
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