摘要
目的 建立DHBsAg免疫细胞化学检测方法 ,研究DHBV肝内原位复制与致病性的关系。方法 采集不同感染状态肝组织标本共 2 2份 ,分别进行DHBsAg免疫组化、Southernblot及组织病理学观察。结果 DHBsAg主要表达于肝细胞胞浆内 ,多呈弥漫性分布 ,无明显胞浆内包涵体形成。急性感染组病毒复制高峰期阳性细胞仅为 2 4 %～ 5 8% ,慢性感染组可 >95 % ;Southern杂交检测急性感染组DHBVDNA为 8.0 5± 2 .72 ,慢性感染组为 18.88± 3.86 (P <0 .0 5 )。不同感染组肝细胞变性坏死、炎症细胞浸润均不明显 ,血清ALT、AST无显著变化。结论 DHBsAg免疫细胞化学方法的建立在细胞水平实现了对病毒表达抗原的定位观察。此外 ,嗜肝病毒的复制不是引起肝细胞损伤的直接原因。
Objective To establish immunocytochemical assay for duck hepatitis surface antigen(DHBsAg)and further analyze the relationship between virus and pathogenesis.Methods Liver samples,taken from acutely and chronically infected ducks,were used to detect DHBsAg by standard immunoperxidase techniques,and DHBV DNA by Southern blot hybridization respectively.Results DHBV surface antigen was usually restricted to randomly scatter cells showing strong cytoplasmic staining.At the peak time of viremia,the positive cells accounted for more than 95% of the total in chronically infected ducks,whereas 24%～58% in acutely infected ones.Moreover,quantitative analysis of DHBV DNA in liver tissue by Southern blot demonstrated significant difference between the two groups(18.88±3.86 vs 8.05±2.72).However,liver histology showed there were no obvious evidence of hepatocyte injury or changes of the level of transaminase during the infected course.Conclusion This methodology will be helpful to illustrate the localization of virus specific antigen in situ.Furthermore,pathogenetic mechanisms for hepatitis were not resulted from replication of virus.
出处
《重庆医学》
CAS
CSCD
2003年第1期51-53,共3页
Chongqing medicine
