摘要
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.
作者
陈倩
陈荫楠
陈东海
林海虹
石贤爱
CHEN Qian;CHEN Yinnan;CHEN Donghai;LIN Haihong;SHI Xian’ai(College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China;Basic Medicine Programme, Quanzhou Medical College, Quanzhou 362000, China;Fujian Key Laboratory of Medical Instrument and Pharmaceutical Technology, Fuzhou 350108, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2017年第20期242-247,共6页
Food Science
基金
国家海洋局海洋公益性行业科研专项(201205022-3)
福建省科技重大专项(2013NZ0003)
福建省海洋与渔业厅重点项目(闽海渔合同[2010]2-27号)
关键词
呋喃唑酮
单链抗体
酶联免疫检测
furazolidone
single chain fragment antibody
enzyme-linked immunosorbent assay