NCM-ELISA检测马铃薯Y病毒(PVY)技术的研究及应用  

Detection of Potato Virus Y by NCM-ELISA Method

作　　者：吕典秋[1] 李学湛[1] 吕文河[2] 于德才[1] 李辉[3] 

机构地区：[1]黑龙江省农业科学院植物脱毒苗木研究所,黑龙江哈尔滨150086 [2]东北农业大学,黑龙江哈尔滨150030 [3]黑龙江省农业科学院谷物品质研究中心,黑龙江哈尔滨150086

出　　处：《中国马铃薯》2006年第6期339-342,共4页Chinese Potato Journal

摘　　要：血清学方法是病毒检测的主要手段。本试验通过对马铃薯Y病毒(PVY)的提纯,免疫家兔制备PVY抗血清,并提取PYV免疫球蛋白IgG作为NCM-ELISA反应为一抗,以市售羊抗兔抗血清为二抗,在硝酸纤维素膜(NCM)上进行NCM-ELISA反应检测PVY,建立PVY NCM-ELISA检测反应体系。试验结果显示,NCM-ELISA具有反应特异性强,灵敏度高的优点,检测植物叶片样品的最高稀释度可达到1:250。通过对田间40份样品的NCM-ELISA和DAS-ELISA检测比较,其检测结果吻合率达到100%。由于NCM-ELISA方法可以将样品点在硝酸纤维素膜上,并且可贮存几个星期或将膜送到其他实验室进行检测,因此具有操作简单,使用方便,检测成本低等优点。Serological technique as important method for viruse diagnose has been applied extensively in the field of disease detection. In this research, potato virus Y （PVY） was purified and the rabbit was immunized with the purified virus of PVY for antiserum preparation. IgG of PVY was prepared as the fist antibody of NCM-ELISA and the commercial antiserum of goat-anti-rabbit was used as the second antibody. The reactions of ELISA were performed on NCM and the formal reaction system was established. The results showed that NCM-ELISA had the same merits of high specificity and sensitivity as DAS-ELISA. The highest detectable dilutions of extract of infected potato leaves was 1：250. NCM-ELISA had 100% consistency with DAS-ELISA based on comparison of test results from 40 potato samples collected from field. The samples can be added onto the NCM and the membrane can be stored for a few weeks or sent to other labs where the test can be done,.so NCM-ELISA has the advantages of easy operation, convenience and low detection cost.

关 键 词：硝酸纤维素膜-酶联免疫分析 马铃薯Y病毒 检测 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]