斑点免疫金染色检测雏鹅新型病毒性肠炎抗体研究

Studies on Dot immunogold staining (IGS) for detecting the antibodies against gosling new type viral enteritis (NGVE)

应用提纯的雏鹅病毒性肠炎病毒（ＮＧＶＥＶ）抗原制备抗原诊断膜，建立了ＩｇＧ的金颗粒标记、抗原抗体反应同步试验的斑点免疫金染色（ＤｏｔＩＧＳ）检测雏鹅新型病毒性肠炎（ＮＧＶＥ，暂定）抗体的方法，只需３０～４０ｍｉｎ即可判断结果；与本动物抗ＧＰＶ，ＤＰＶ，ＮＤＶ，ＩＢＶ，ＩＬＴＶ，ＭＤＶ，ＩＢＤＶ，ＥＤＳＶ及多杀性巴氏杆菌血清不发生交叉反应；对兔抗ＮＧＶＥＶ的ＩｇＧ的最低检出量为１．０×１０－１０ｇ／ｍＬ。雏鹅感染ＮＧＶＥＶ强毒后第３ｄ、成年鹅接种ＮＧＶＥＶＣＮ４０弱毒疫苗后第３ｄ即可检测到相应抗体。对重庆市和四川省１０个县（市）的６１７份成年鹅血清进行检测，结果ＮＧＶＥ的血清阳性率为３０．４４％～３６．８４％。该法适用于雏鹅感染的早期诊断、成年鹅的血清流行病学调查和疫苗免疫效果评价等。

The purified gosling new type viral enteritis viruses (NGVEV) were used to prepare the antigen diagnostic membrane and to establish the Dot immunogold staining method of IgG gold particle labbeled antigen antibody synchronous test for detecting NGVE antibodies, by which an accurate result diagnosing NGVE would be obtained within 30 to 40 min. The method has a better specificity but no cross reaction with positive sera of gosling plague, duck plague, newcastle disease, avian infectious bronchitis, infectious larynogtracheitis, Marek's disease, infectious bursal disease, egg drop syndrome and avian pasteurellosis. And by this method the minimum amount of rabbit anti NGVEV IgG as low as 1.0×10 10 g/mL can be detected. The antibodies could be detcted on the third day after both goslings were infected with virulent NGVEV and adult gooses were inoculated with NGVE attenuated vaccine. 617 serum samples of adult gooses from ten districts of Sichuan province and Chong Qing city, and the positive detectable rates were 30.44 to 36.84 percent. The method is suitable for early diagnosing NGVE in goslings and investigating seroepidemiologically NGVE in adult gooses and evaluating the efficacies of the vaccination.