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当前位置: 首页 > 文章 > 牙鲆淋巴囊肿病毒双抗体夹心ELISA检测方法的建立 水产科学 2009,28 (7) 374-377
Position: Home > Articles > Development of Double-antibody Sandwich Indirect ELISA to Detect Lymphocystis Disease Virus in Japanese Flounder Fisheries Science 2009,28 (7) 374-377

牙鲆淋巴囊肿病毒双抗体夹心ELISA检测方法的建立

作  者：
程顺峰
单  位：
青岛农业大学动物科技学院
关键词：
淋巴囊肿病毒;ELISA;建立;双抗体夹心
摘  要：
以牙鲆淋巴囊肿病毒(Lymphocystis disease virus,LCDV)特异性单克隆抗体为捕获抗体,纯化的兔抗LCDV血清为检测抗体,建立了LCDV病毒双抗夹心间接ELISA检测方法。该方法最佳反应条件为:单克隆抗体包被质量浓度为2μg/ml,兔抗血清工作浓度为1∶1200稀释,以3%牛血清白蛋白作为封闭液。应用本方法对体表具有明显发病症状的牙鲆、与发病鱼同池但体表无症状的牙鲆及健康牙鲆的肝、脾、肾、胃、肠、心脏、鳃等7种组织进行检测,在前两种鱼的胃、肠、鳃等组织检测到病毒,而健康牙鲆各组织器官反应结果均为阴性,试验结果表明,所建立的ELISA检测方法具有特异性强、可靠性高的特点,可用于养殖牙鲆LCDV的检测。
译  名：
Development of Double-antibody Sandwich Indirect ELISA to Detect Lymphocystis Disease Virus in Japanese Flounder
作  者：
CHENG Shun-feng(College of Animal Science and Technology,Qingdao Agricultural University,Qingdao 266109,China)
关键词：
lymphocystis disease virus;sandwich indirect ELISA;development
摘  要：
A sandwich enzyme linked immunosorbent assay(ELISA) was developed to detect lymphocystis disease virus(LCDV) in Japanese flounder(Paralichthys olivaceus) by monoclonal antibody against LCDV as capture antibody and purified rabbit as detector antibody.The 96-well enzyme immunoassay(EIA) plates were coated with 100 μl monoclonal antibody at concentration of 2 μg/ml and blocked with 0.3% bovine serum albumin(BSA).The working concentration of rabbit IgG purified from polyclonal antibody was diluted 1∶1200 with distilled water.The ELISA method was applied to detect the LCDV in the stomach,intestine,liver,spleen,kidney and gill in the flounder with lymphocystis nodules on body surface,the flounder without obvious symptoms cultured in the same tank,and healthy flounder.The results showed that the stomach,intestine and gill in the flounder with lymphocystis nodules on body surface were infected with LCDV.In addition,the flounder without obvious symptoms cultured in the same tank were also detected virus and the locations of signal in tissues were the same with that seriously.However,for the healthy flounder,no LCDV infection was found by the ELISA method,indicating that this method can be used to diagnose lymphocystis disease in Japanese founder efficiently,especially the ones without obvious symptoms.

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