摘要
以蔗糖密度梯度离心提纯的Ⅰ型鸭肝炎病毒(DHV-Ⅰ)免疫兔制备兔抗DHV-Ⅰ抗体,建立了检测石蜡组织切片中DHV-Ⅰ抗原的间接免疫酶染色(indirect immunoperoxidase staining,IIS)方法,并对DHV-Ⅰ强毒人工感染死亡或濒死雏鸭的各个组织器官进行了检测。结果表明,IIS与DHV-Ⅰ感染死亡或濒死雏鸭的肝等呈现阳性反应,与鸭瘟病毒、鸭疫里默氏杆菌、大肠杆菌、沙门菌、多杀性巴氏杆菌感染发病和死亡雏鸭的肝以及健康雏鸭的肝呈现阴性反应。感染DHV-Ⅰ死亡或濒死雏鸭的肝、脾、肾、心、胸腺、腔上囊、胰腺、十二指肠、盲肠、空肠、回肠、直肠的免疫组织化学检测呈阳性或强阳性,DHV-Ⅰ抗原主要分布于感染细胞的细胞质。该IIS法具有良好的特异性,可用于DHV-Ⅰ在感染雏鸭组织细胞中的亚细胞定位、DHV-Ⅰ感染雏鸭的实验室诊断、甲醛固定组织的回顾性诊断。
To detect duck hepatitis virus type Ⅰ （DHV-Ⅰ ） and to localize virus antigens in infected cells using an immunoperoxidase staining method,antibodies against DHV-Ⅰ were prepared from the rabbits vaccinated with purified DHV-Ⅰ by sucrose gradient centrifugation. An indirect immunoperoxidase staining（ⅡS） test was established for the detection of DHV-Ⅰ antigen in the paraffin wax tissue splice. DHV-Ⅰ in each organ tissue of dead or dying ducklings experimentally infected with DHV-Ⅰ were detected. Using ⅡS,the positive reaction was observed in liver and other tissues of dead or dying ducklings infected with DHV- Ⅰ ,whereas negative reaction was observed in the liver of dead ducklings infected respectively with duck plague virus, duck＇s Riemerella anatipestifer, Escherichia coli, Salmonella and Pasteurella multocida and the livers of healthy ducklings. The liver, spleen, kidneys, heart, thymus, pancreas, duodenum,jejunum,ileum,cecum and rectum from dead or dying ducklings experimentally infected with DHV- Ⅰ ,were positive or strongly positive by ⅡS. The DHV- Ⅰantigen was mainly distributed in the cytoplasm of infected cells. The established IIS was specific, and could be applied for the subcellular localization of DHV- Ⅰ antigen in tissue cell of infected ducklings. It can be used for experimental diagnosis of DHV- Ⅰ infection in duckling and for review diagnosis of formaldehyde fixed tissue.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第5期369-373,共5页
Chinese Veterinary Science
基金
国家科技攻关重大项目(2004BA901A03)
教育部新世纪优秀人才支持计划项目(NCET-04-0906)
四川省基础研究重大项目(05JY029-109)
四川省重点建设学科项目(SZD0418)
关键词
免疫组织化学
检测
鸭肝炎病毒
鸭
抗原定位
immunohistochemistry
detection
duck hepatitis virus
duck
localization antigen
