Alexandrov M, Alexandrova R, Alexandrov I, Zacharieva S, Lasarova S, Doumanova L, Peshev R, Donev T
Institute of Experimental Pathology and Parasitology, Bulgarian Academy of Sciences, Sofia.
J Virol Methods. 1999 Apr;79(1):21-32. doi: 10.1016/s0166-0934(98)00175-x.
Polyclonal antibodies (PAbs) raised in geese and eight mice hybridomas secreting monoclonal antibodies (MAbs) against the goose parvovirus (GPV) were prepared. They were used for development of immunofluorescence (IF) and immunoelectron-microscopic (IEM) techniques to demonstrate the GPV infection in infected organs and biological fluids. The GPV antigens were established by immunofluorescence within the nuclei and the cytoplasm of many infected cells of the chorioallantoic membrane of goose and Peckin duck embryos, liver and heart of mortally diseased goslings. By means of IEM it was possible to detect the GPV in native organ homogenate supernatants and allantoic fluids. All techniques used in the study could be successfully applied for rapid diagnosis of the GPV infection. The test systems on the basis of MAbs should, however, be preferred. By means of immunoblotting (IB) using PAbs and MAbs four viral proteins (VP) with MW 88, 77, 65 and 60 kDa were demonstrated. Contrary to the others the VP with MW 65 kDa was the most antigenically reactive though invisible in the SDS-PAGE and Coomassie-blue dye-stained preparations.
制备了在鹅体内产生的多克隆抗体（PAbs）以及8株分泌抗鹅细小病毒（GPV）单克隆抗体（MAbs）的小鼠杂交瘤。它们被用于开发免疫荧光（IF）和免疫电子显微镜（IEM）技术，以证明在受感染器官和生物体液中的GPV感染。通过免疫荧光在鹅和北京鸭胚胎的绒毛尿囊膜、病死雏鹅的肝脏和心脏的许多受感染细胞的细胞核和细胞质中确定了GPV抗原。借助IEM能够在天然器官匀浆上清液和尿囊液中检测到GPV。该研究中使用的所有技术都可成功应用于GPV感染的快速诊断。然而，基于MAbs的检测系统应更受青睐。使用PAbs和MAbs通过免疫印迹（IB）证明了4种分子量分别为88、77、65和60 kDa的病毒蛋白（VP）。与其他蛋白不同，分子量为65 kDa的VP具有最强的抗原反应性，尽管在SDS-PAGE和考马斯亮蓝染色制剂中不可见。
