抗柱状黄杆菌多克隆抗体的制备及其免疫学特性鉴定

为制备抗柱状黄杆菌多克隆抗体,本研究利用颗粒性抗原免疫新西兰白兔,收集的抗血清通过辛酸-硫酸铵法纯化,采用间接ELISA法检测纯化后多克隆抗体的效价和交叉反应性。结果显示,制备的多克隆抗体蛋白质浓度为29.28 mg/mL,效价在1:6.4×10⁴以上,与迟钝爱德华氏菌、大肠杆菌、嗜水气单胞菌、鳗弧菌、溶藻弧菌、副溶血弧菌及哈维氏弧菌等水生动物致病菌均无交叉反应。本研究成功建立了抗柱状黄杆菌多克隆抗体的制备方法,可用于柱状黄杆菌的快速检测。

Preparation of the Polyclonal Antibody against Flavobacterium columnaris and Identification of its Immunological Properties

To prepare the polyclonal antibody against Flavobacterium columnaris, New Zealand rabbits were immunized with particulate antigen. Antisera from the immunized rabbits were purified with caprylic/ammonium sulfate precipitation. Titers and cross reaction of the polyclonal antibody were determined by indirect ELISA. The results showed that the protein concentration of the polyclonal antibody was 29.28 mg/mL. The titer was over 1:6.4×10⁴. No cross-reaction was observed with other aquatic pathogenic bacteria such as Edwardsiella tarda, E.coli, Aeromonas hydrophila, Vibrio anguillarum, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio harveyi. High specific polyclonal antibody against Flavobacterium columnaris was prepared, which might be used for rapid detection of Flavobacterium columnaris.