抗绿脓杆菌外毒素A酶标抗体的制备及其应用

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0.0608 mg/mL和0.336 mg/mL;HRP/Ab(IgG)克分子比值为1.724%;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、特异性强的优点.

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

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.