摘要
为建立一种检测Ⅰ群禽腺病毒(FAVⅠ)抗体的间接ELISA方法,以FAVⅠhexon重组蛋白作为包被抗原,优化反应条件,建立了FAVⅠ抗体间接hexon-ELISA检测方法。抗原最佳包被浓度为10.0μg/mL,最适包被条件为37℃2h加4℃过夜;最佳封闭液为50g/L脱脂乳;血清最佳稀释度为1∶100,最佳作用时间为75min;酶标二抗最佳工作浓度为1∶2 000,最佳作用时间为45min;hexon-ELISA阴性、阳性临界值为0.379。用建立的hexon-ELISA方法检测100份鸡血清样品,阳性率为45%。结果表明,建立的hexon-ELISA方法简便、快捷,可大批量检测,具有良好的特异性、敏感性和重复性,适用于FAVⅠ的诊断、抗体水平监测及流行病学调查。
An indirect enzyme-linked immunosorbent assay（ELISA） method was developed to detect antibody against fowl adenovirus group Ⅰ （FAV Ⅰ ）. The recombinant protein hexon of FAV Ⅰ was used as coating antigen for the ELISA method,and the reaction conditions were optimized. The optimal concentra- tion of antigen was 10.0 μg/mL and the coating condition was 37 ℃ 2 h and 4℃ overnight. The optimal blocking buffer was 50 g/L skimmed milk,the optimal dilution and reaction time of serum was 1 ： 100 and 75 min,respectively;and the optimal dilution and reaction time of enzyme-labeled antibody were 1：2 000 and 45 min, respectively. The critical value of the hexon-ELISA was 0. 379. Furthermore, one hundred chicken serum samples were detected by the hexon-ELISA, and the positive rate of FAV I antibody was found to be 45%. The results suggested that the hexon-ELISA is a specific, sensitive and reliable method for the diagnosis, antibody level surveillance and epidemiologic investigation of FAV Ⅰ.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2012年第7期701-707,共7页
Chinese Veterinary Science
基金
广西特聘专家专项经费资助项目(2011B020)
广西科技项目(桂科攻0815009-3-6
2010GXNSFA013090
2010GXNSFA013089)
