摘要
表达并纯化了禽呼肠孤病毒(ARV)的2个非结构蛋白σNS和P17,并以此作为包被用抗原,分别进行间接ELISA。结果表明,抗原最佳包被浓度分别为9.3μg/mL和11.5μg/mL;一抗血清的稀释度都是1∶200;HRP酶标二抗羊抗鸡IgG稀释度为1∶5 000,初步建立了能检测ARV感染的σNS-ELISA、P17-ELISA方法。以σNS、P17两种蛋白按以上确定的条件同时包被,建立了σNS-P17-ELISA。分别用σNS-ELISA、P17-ELISA及σNS-P17-ELISA对接种过ARV活病毒或灭活疫苗的33份SPF鸡血清进行检测,结果发现以非结构蛋白建立的3种ELISA方法均能区分ARV活病毒感染与灭活疫苗免疫的抗体,进一步对这3种方法进行敏感性、特异性分析比较,表明σNS-P17-ELISA方法能更有效地区分ARV感染与灭活疫苗免疫抗体。
The σNS and P17 fusion proteins of avian reovirus （ARV） were expressed and purified, σNS-ELISA and P17-ELISA were developed by using the σNS and P17 fusion proteins as the coating antigen, respectively. The optimal concentrations of σNS and P17 fusion proteins were 9. 3 μg/mL and 11.5 μg/mL, respectively, the dilution of serum sample was 1 ： 200, and the optimal dilution of enzyme-labeled goat anti-chicken IgG was 1 ： 5 000. The σNS-P17-ELISA was developed by using the purified σS and P17 fusion proteins together as the coating antigen. Thirty-three sera from SPF chickens, which were infected with ARV or vaccinated with ARV inactived vaccine, were detected by the above three ELISA. The results showed that the three ELISA could distinguish between the vaccinated and infected chickens, and the σNS-P17-ELISA was the most sensitive and specific.
出处
《畜牧与兽医》
北大核心
2009年第11期11-15,共5页
Animal Husbandry & Veterinary Medicine
基金
国家百千万人才工程人选专项资金项目(945200603)
广西留学回国人员基金项目(桂科回0832019
0639016)
广西自然科学基金项目(桂科自0991222)。
