抗绿脓杆菌外毒素A酶标抗体的制备及其应用  

The preparation and application of the pseudomonas aeruginosa Exotoxin IgG-AP

作　　者：高玲美[1] 牛钟相[1] 李雅林[1] 丁淑燕[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国预防兽医学报》2005年第1期67-70,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：山东省科技厅"三○工程"项目研究内容

摘　　要：从病死羊体内分离到绿脓杆菌并提取出外毒素A(PEA) ,再以此毒素作为抗原加油佐剂制成乳化抗原免疫家兔 ,获得高免血清并提取免疫球蛋白G(IgG) ;用过碘酸钠法将过氧化物酶标记抗PEA抗体 (IgG) ,制成酶标抗体 ,经检验 ,酶标抗体结合物中的HRP浓度和Ab(IgG)浓度分别是 0 0 6 0 8mg/mL和 0 336mg/mL ;HRP/Ab(IgG)克分子比值为1 72 4 % ;酶 (HRP)结合率是 11 15 %。利用该酶标抗体以ELISA夹心法对羊体内抗PEA抗体含量进行了检测 ,结果证明 ,用酶标抗体ELISA法比用平板凝集实验法检测的抗体效价平均高出二个滴度 ,表明制备的PEA酶标抗体具有灵敏度高。Pseudomonas aeruginosa was isolated from sick sheep then Pseudomonas Exotoxin A(PEA) was extracted from Pseudomonas aeruginosa.The toxin was used as antigen with oil-adjuvant to make into oil-emulsified vaccine to immune rabbits.High-immunity serum was got and IgG was extracted from the serum;With the method of NaIO-4 anti-PEA antibody was labelled to make enzyme-labelled antibody.After tested,in the combination of enzyme-labelled antibody,the concentration of HRP and Ab(IgG) was (0.0608?mg/mL) and 0.336?mg/mL respectively;HRP/Ab(IgG) was 1.724?%;the combination rate of enzyme was 11.Using the enzyme-labelled antibody,the concentration of anti-PEA antibody was tested in the sheep with the method of sandwich ELISA.It proved that the antibody valence that was tested with the method of enzyme-labelled ELISA was 2 valence higher than that of with the method of flat-plate agglutination experiment.So the PEA enzyme-labelled antibody has the advantage of high sensitivity and specificity.

关 键 词：绿脓杆菌 外毒素A 酶标抗体 

分 类 号：S852.61[农业科学—基础兽医学]