论文部分内容阅读
为建立一种通过胶体金免疫层析技术快速检测柱状黄杆菌的方法,试验采用柠檬酸三钠还原法制备粒径为20nm的胶体金颗粒,将其标记纯化的抗柱状黄杆菌单克隆抗体(McAb)制备出金标抗体结合垫。纯化的兔抗柱状黄杆菌多克隆抗体(PcAb)和羊抗鼠IgG分别包被在硝酸纤维素膜的检测线(T)与质控线(C)上,制备出胶体金免疫层析试纸条,并对试纸条的灵敏度、特异性及稳定性进行测定。结果显示,该试纸条检测灵敏度为1×103 CFU,检测时间为3.5min,制备的试纸条与迟钝爱德华氏菌、大肠杆菌、嗜水气单胞菌、鳗弧菌、溶藻弧菌、副溶血弧菌、哈维氏弧菌均无交叉反应,且稳定性好。本研究首次成功建立了柱状黄杆菌胶体金快速检测方法,所制备的试纸条具有灵敏、特异、稳定、快速等优点,可用于柱状黄杆菌的检测。
In order to establish a rapid detection method of Flavobacterium columnarum by colloidal gold immunochromatography, colloidal gold particles with a particle size of 20nm were prepared by the method of trisodium citrate reduction and labeled with the purified monoclonal antibodies against L. cylindromator McAb) gold standard antibody conjugate pad. The purified rabbit polyclonal anti-bacterium IgG (PcAb) and goat anti-mouse IgG were respectively coated on the detection line (T) and the control line (C) of the nitrocellulose membrane to prepare a colloidal gold immunochromatographic test strip Article, and test strip sensitivity, specificity and stability were determined. The results showed that the test strip test sensitivity of 1 × 103 CFU, the test time was 3.5min, prepared test strips and the retarded Edwardsiella, Escherichia coli, Aeromonas hydrophila, Vibrio anguillarum, Vibrio alginolyticus , Vibrio parahaemolyticus, Vibrio harveyi no cross-reaction, and good stability. In this study, for the first time, the rapid detection method of Flavobacterium columnarum was successfully established. The prepared strip has the advantages of sensitivity, specificity, stability and rapidity and can be used for the detection of Flavobacterium columnarum.