应用单克隆抗体检测大麦黄花叶病毒

THE DETECTION OF BARLEY YELLOW MOSAIC VIRUS WITH MONOCLONAL ANTIBODY OF BaYMV

作者应用已建立的大麦黄花叶病毒(BaYMV)的4F_(10)单克隆抗体杂交瘤细胞株,经小白鼠腹水生产单克隆抗体,用过碘酸钠法制备了辣根过氧化物酶标记的酶标单克隆抗体。并由此建立了检测大麦黄花叶病毒的标准的双抗体夹心酶联免疫吸附法(DAS-ELISA)。其检测提纯病毒的灵敏度为3.0μg/ml左右,感病大麦叶片汁液的最大稀释度为1:2560,病茎稀释度为1:160。对采自我国7个主要BaYMV发病区感病的样品进行了检测,绝大多数均显示强阳性,只有宝山样品例外。本项试验为BaYMV诊断的标准试剂盒的建立进行了有益的尝试。

The McAbs secreted by 4F10 hybridoma cell line against BaYMV were purified from ascitic fluids. The HRP was conjugated to the McAbs through periiodate coupling methods. A direct DAS-ELISA method for detecting BaYMV was developed by using the McAb-HRP. Results indicated that the minimum amount of purified BaYMV could be detected as little as 3. 0*g/ml and the BaYMV could be detected in leaves and stems of infected barley plants at dilutions of 1/2560 and 1/160 respectively by this method. Seven samples of plants infected with BaYMV collected from different severely diseased locations in China winter barley area were detected by this method,and the results showed that six samples reacted positively with the McAb, but the sample from Baoshan reacted negatively.