【作者】 查智辉； 施慧； 谢建军； 许文军；

【Author】 ZHAZhi-hui~(1,2),SHI Hui~2,XIE Jian-jun~2,XU Wen-jun~(1,2) (1.Marine and Fishery Research Institute of Zhejiang Ocean University,Marine and Fisheries Research Institute of Zhejiang Province,Zhoushan,Zhejiang 316100.China;2.Marine Science School of Zhejiang Ocean University.Zhoushan,Zhejiang 316100,China)

【机构】 浙江省海样水产研究所； 浙江海洋学院海洋与渔业研究所； 浙江海洋学院海洋科学学院；

【摘要】 目的制备抗血卵涡鞭虫的单克隆抗体(McAb)并研究其初步应用。方法用血卵涡鞭虫可溶性抗原免疫8周龄雌性BALB/c小鼠,经常规融合,以间接ELISA方法筛选出分泌高滴度、高特异性单克隆抗体的杂交瘤细胞株,并使用所得单抗检测血卵涡鞭虫样本,最终建立双抗体夹心ELISA法检测感染血卵涡鞭虫的三疣梭子蟹。结果获得3株分泌高滴度抗血卵涡鞭虫可溶性抗原的单克隆抗体杂交瘤细胞株(282、3G4、4G7),分泌抗体经鉴定均为IgG。双抗体夹心ELISA法和间接荧光抗体试验结果表明,三株单抗都与血卵涡鞭虫可溶性抗原呈强阳性反应,而不与三疣梭子蟹其它病原孢子虫、溶藻弧菌及三疣梭子蟹血淋巴等发生交叉反应。双抗体夹心ELISA检测血卵涡鞭虫可溶性蛋白抗原灵敏度为100pg/ml。结论制备的抗血卵涡鞭虫可溶性抗原的杂交瘤细胞株能分泌高滴度、高特异性的单克隆抗体,且在病原的临床检测方面有应用前景。更多还原

【Abstract】 Objective To develop and identify monoclonal antibodies(McAbs) against Hematodinium sp.in Portunus trituberculatus.and observe its applicability.Methods 8-week-old female BALB/c mice were immunized with soluble antigen of Hematodinium sp..The:spleen cells of immunized mice were fused with myeloma cell,and the hybridoma secreting high titer of McAbs with high specificity was screened.By using the McAbs,Hematodinium sp.was detected by indirect ELISA.Double antibody sandwich enzyme-linked immunosorbent assay(ELISA) respectively.Results Three McAbs were established (2B2,3G4,4G7),which all belonged to the IgG subtype.Three clones all showed no cross reaction with antigens of Nosema,V.alginolyticus and the haemolymph of Portunus trituberculatus.The sensitivity of the double antibody sandwich ELISA to detect Hematodinium sp.antigen was 100pg/ml.Conclusion Three hybridoma cell lines against Hematodinium sp.in Portunus trituberculatus have been established which secret high titer of McAbs with high specificity and seem promising in detecting the Hematodinium sp.in Portunus trituberculatus.更多还原