左氧氟沙星人工抗原间接ELISA检测方法的建立  

Establishing the Method for Detecting Levofloxacin Artificial Allergens by Indirect Enzyme-linked Immunosorbent Assay

作　　者：张冠世[1] 张凤凤[1] 胡亚南[1] 肖佳佳[1] 裴凯祥[1] 李林育[1] 黄瑞蕊[1] 李引乾[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《西北农业学报》2012年第3期51-54,70,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：西北农林科技大学国家级大学生创新项目

摘　　要：为检测所制备的左氧氟沙星人工抗原品质,采用西北农林科技大学兽医药理实验室制备的左氧氟沙星人工抗原免疫BALB/c小鼠,制得鼠抗左氧氟沙星人工抗原的多克隆抗体。以采集的抗血清为一抗、辣根过氧化物酶标记的羊抗鼠IgG为二抗、四甲基联苯胺(TMB)为底物,建立左氧氟沙星人工抗原间接酶联免疫检测法(ELISA)。试验结果表明,间接ELISA最佳反应条件为包被抗原质量浓度1mg/L、4℃过夜、抗原抗体于37℃作用1h、酶标二抗稀释度为1∶2 000。该方法的灵敏度高,且具有良好的准确性和重复性,可以评价左氧氟沙星人工抗原的品质。BALB/c mice were immunized by artificial allergens of levofloxacin made in our laboratory in order to detect its quality and obtain polyclonal antibody.Taken collected serum as first antibody,goat anti-mouse IgG labeled by horseradish peroxidase（HRP） as secondary antibody,tetramethylbenzidine（TMB） as substrate,indirect enzyme-linked immunosorbent assay（ELISA） was established.The results showed that the optimal conditions are as follows： coating antigen concentration was 1 mg/L,coated overnight at 4 ℃,reaction condition between antigen and antibody was 1 h at 37 ℃,and the dilution of HRP secondary antibody was 1∶2 000.This method has a good sensitivity,accuracy and repeatability for the detection of LVL artificial allergens.

关 键 词：左氧氟沙星 人工抗原 间接ELISA 检测 

分 类 号：S859.84[农业科学—临床兽医学]