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:抗原包被浓度50mg/L,37℃作用2h后4℃过夜包被,血清的稀释度为1∶320,抗原抗体反应时间为50min,酶标二抗稀释度为1∶12 000,作用时间为30min,底物显色时间为15min。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。
An indirect ELISA coated with supernatant of sonicated Haemophilus parasuis serovar5 cell culture was established.The results of chess board titration tests showed that coating antigen at a mass concentration of 50 mg/L incubated optimally at 37 ℃ for 2hand then coated overnight at4 ℃.The optimal dilution of serum and HRP-IgG was 1∶320and 1∶12 000,respectively.The reaction time of coating antigen with serum and serum with HRP-IgG 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.
出处
《西北农业学报》
CAS
CSCD
北大核心
2015年第5期29-33,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
2014年地方高校国家级大学生创新创业训练计划(201410467022)
新乡市重点科技攻关(ZG13009)
2014年河南科技学院大学生创新项目
