对虾白斑综合征病毒单抗介导间接ELISA的建立

从白斑综合征病毒（WSSV）青岛株感染的克氏原螯虾（Canbarus proclarkii）中提纯病毒，用纯化病毒免疫BALB／c小鼠，采用细胞融合法获得4株阳性杂交瘤细胞，分别命名为1B1、1E4、4E6和4E5。4株单抗均为IgM。4E5株单抗在免疫转印中与37500左右的病毒蛋白条带呈阳性反应。用此株单抗作一抗，建立检测病毒蛋白的间接ELISA。该方法用于人工感染WSSV的螯虾组织样品中病毒的检测，48h后即有阳性检出，而正常螯虾组织均呈阴性。

Establishment of Monoclonal Antibody Mediated Indirect ELISA for Detection of White Spot Syndrome Virus(WSSV) and It＇s Application in Crawfishes

White spot syndrome virus(WSSV) was purified from infected crawfishes (Cambarus proclarkii).The purified virus was used as antigen to immunize BALB/c mice and four monoclonal antibodies(McAbs),designated as 1B 1,1E 4,4E 6 and 4E 5,were obtained by hybridoma technique.The four McAbs all belong to IgM.One of them(4E 5),reacted with an approximately 37 500 viral protein band in Western blotting analysis,was selected for establishment of McAb-mediated indirect ELISA.The ELISA was applied to detect WSSV in crawfishes.The first positive reaction was at 2 day post inoculation and all reactions were positive at 5 day post inoculation in infected crawfishes,while all reactions were negative in healthy crawfishes.This ELISA was proved to be effective and convenient for detection of WSSV in crawfishes.