ELISA双抗体夹心法检测河蟹“颤抖病”病毒  

Detection of Eriocheir sinensis (ES)"tremble disease"virus by double antibody sandwich enzyme linked immunosorbent assay(ELISA)

作　　者：孙学强[1] 金业[1] 陆承平[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,南京210095

出　　处：《南京农业大学学报》2001年第4期67-70,共4页Journal of Nanjing Agricultural University

基　　金：江苏省科委应用基础项目 (Q990 2 )

摘　　要：用纯化的河蟹“颤抖病” (暂定名 )病毒分别免疫家兔和山羊 ,制备高效价的抗血清。羊抗血清经硫酸铵盐析纯化。经方阵滴定 ,选择P/N =2 95时作 1∶10 0稀释的抗血清为工作浓度 ,建立ELISA双抗体夹心法检测样本 14 4份 ,包括来自长江水域的野生河蟹、人工感染发病的河蟹、 1999年和 2 0 0 0年从江苏采集的自然发病的病蟹及市场购买河蟹。结果显示 ,病蟹和市场购买河蟹的阳性率为 6 6 7%～ 10 0 % ,人工感染的病蟹和人工感染石蟹致病组阳性率均为 10 0 % ,来自长江水域的健康河蟹及未接种病毒的对照石蟹阳性率均为 0 ,表明建立的方法可以用于检测河蟹“颤抖病”The antisera against purified Eriocheir sinensis'tremble disease'virus was abtained from the rabbits and goat.The optimal dilution of antisera with positive/negative 2 95 was determined as 1∶100.After the establishment of double antibody sandwich ELISA,144 samples from the Yangtze River,markets and experimental infected or natural dieased in Jiangsu Province were detected.The results showed that the P/N of the ES from the exposed region and markets was 66 7%～100%.The experimental infected ES,inoculated with purified virus or the supernatant of the diseased ES homogenate were all positive,but the wild healthy ES from the Yangtze River and the controls were all negative.It suggested that the established ELISA could be used to detect the virus of the Eriocheir sinensis suffered'tremble disease'.

关 键 词：河蟹 ELISA双抗体夹心法 “颤抖病”病毒 检测 

分 类 号：S947.9[农业科学—水产养殖]