【作者】 马福利； 王丽荣； 董永军； 刘运平； 王秋霞； 梁莹莹；

【Author】 MA Fuli;WANG Lirong;DONG Yongjun;LIU Yunping;WANG Qiuxia;LIANG Yingying;College of Animal Science,Henan Institute of Science and Technology;Antibody Engineering Laboratory,Henan Institute of Science and Technology;Xinxiang Generalization Station of Animal Husbandry Technology;

【机构】 河南科技学院动物科学学院； 河南科技学院抗体工程重点实验室； 新乡市畜牧技术推广站；

【摘要】 采用超声裂解副猪嗜血杆菌分离株5型菌体上清作为包被抗原,初步建立副猪嗜血杆菌抗体的间接ELISA检测方法。通过棋盘滴定法筛选确定最佳反应条件:抗原包被质量浓度50 mg/L,37℃作用2 h后4℃过夜包被,血清的稀释度为1:320,抗原抗体反应时间为50 min,酶标二抗稀释度为1:12 000,作用时间为30 min,底物显色时间为15 min。经特异性、敏感性和符合性试验检测结果表明,建立的间接ELISA方法特异性和重复性良好,敏感性高,可用于副猪嗜血杆菌的检疫和流行病学调查。更多还原

【Abstract】 An indirect ELISA coated with supernatant of sonicated Haemophilus parasuis serovar 5cell culture was established.The results of chess board titration tests showed that coating antigen at a mass concentration of 50mg/L incubated optimally at 37 ℃ for 2 h and then coated overnight at 4℃.The optimal dilution of serum and HRP-Ig G was 1:320 and 1:12 000,respectively.The reaction time of coating antigen with serum and serum with HRP-Ig G was 50 min.The time of color development reaction was 15 min.Specificity,sensitivity and compliance tests confirmed that the developed ELISA had good stability and specificity.So this assay can be used as a tool for diagnosis and quarantine of Haemophilμs parasμis.更多还原