普多沙星抗体的制备及其在兽药残留检测中的初步应用

Preparation of Pradofloxacin Antibody and Its Preliminary Application in Detection of Veterinary Drug Residues

普多沙星(Pradofloxacin)是一种动物专用的新型第3代氟喹诺酮类抗菌药。普多沙星虽然为犬猫专用药物,但仍存在将其用于食品动物的可能,而动物源食品中兽药残留会损害人类的健康。因此,有必要建立快速高效的普多沙星残留检测方法。酶联免疫吸附测定法(ELISA)非常适合于抗生素药物残留的快速检测。该研究利用化学偶联方法将普多沙星与牛血清白蛋白(BSA)和鸡卵清蛋白(OVA)成功偶联,获得了普多沙星-BSA和普多沙星-OVA,并利用紫外—可见分光光度法对合成的免疫原进行初步鉴定分析。以人工抗原普多沙星-BSA为免疫原免疫实验兔,制备抗血清,最终建立了检测普多沙星的间接竞争ELISA方法。本方法的检测限0.0238μg/mL,检测范围为0.0238~4.010μg/mL。

Pradofloxacin is a novel animal-specific third-generation fluoroquinolone.Although pardofloxacin is only approved for dogs and cats now,there is still risk of use in food-producing animals,and the residues of veterinary drugs in animal-origin foods will harm human health.Therefore,it is necessary to establish a rapid and efficient method for the detection of pardofloxacin residues.Enzyme-linked immunosorbent assay(ELISA)is very suitable for rapid detection of antibiotic drug residues.In this study,a chemical coupling method was used to successfully couple pradofloxacin with bovine serum albumin(BSA)and chicken ovalbumin(OVA)to obtain the pradofloxacin-BSA and pradofloxacin-OVA.The UV-Vis spectrophotometry was used to identify and analyze the synthesized immunogen.The experimental rabbits were immunized with artificial antigen,pradofloxacin-BSA,to prepare antiserum.Finally,an indirect competitive ELISA method for the detection of pradofloxacin was established.The detection limit of this method was 0.0238μg/mL and the detection range was 0.0238~4.010μg/mL.