基于单链抗体的呋喃唑酮酶联免疫检测方法的建立

Establishment of Enzyme-Linked Immunosorbent Assay Method for Detecting Furazolidone Based on Single Chain Fragment Antibody

目的:为快速检测呋喃唑酮(furazolidone,FZD)在动物性食品中的残留量。方法:基于单链抗体的间接竞争酶联免疫吸附实验法建立了FZD检测方法。结果:最佳抗原工作质量浓度为2μg/m L,最佳抗体稀释倍数为1∶500,一抗最佳反应时间为60 min,二抗最佳反应时间为45 min,四甲基联苯胺最佳显色时间为20 min。FZD检测试剂盒在10~100 ng/m L范围具有较好的线性关系,IC50值为13.01 ng/m L,检出限为1.28 ng/m L,回收率为73.38%~84.52%。结论:与抗FZD单克隆抗体相比,所建立的检测试剂盒检测范围更广,且具有很高的灵敏度以及很好的特异性和稳定性。

Aim:This study aimed to develop an enzyme-linked immunosorbent assay(ELISA)method for detecting the residues of furazolidone(FZD)in animal food.Method:The indirect competitive ELISA method based on single chain fragment antibody was established.Result:The optimal antigen mass concentration was2μg/mL,and the optimal antibody dilution ratio was1:500,and the optimal reaction time of primary antibody,the optimal reaction time of secondary antibody and the optimal reaction time of TMB were60min,45min,and20min,respectively.The good linearity was seen in the range of10-100ng/mL of FZD,with the IC50value being13.01ng/mL,and the lowest detection limit(LOD)being1.28ng/mL,and the recovery rates being73.38%-84.52%.Conclusion:Compared with the monoclonal antibody against FZD,the detection kit based on single chain fragment antibody displayed wider detection range,higher sensitivity,better specificity and detection stability.