3种方法检测黄瓜绿斑驳花叶病毒的灵敏度对比分析  

Comparison of Three Techniques for Detection the Sensitivity of Cucumber Green Mottle Mosaic Virus

作　　者：李海明[1,2] 沈建国[3] 吴祖建[1] 陈启建[1] 

机构地区：[1]福建农林大学植物病毒研究所,福州350002 [2]福建省农科院甘蔗研究所,福建漳州363005 [3]福建出入境检验检疫局,福州350001

出　　处：《中国农学通报》2010年第17期269-272,共4页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金项目"鸦胆子素D抗烟草花叶病毒机制研究"(30770089);福建省科技重大专项"桔小实蝇和红火蚁的预警与监测技术研究"(2006NZ0002-2);福建出入境检验检疫局科技项目"进境果蔬上黄瓜绿斑驳花叶病毒分子检测技术的研究"(FK2008-05)

摘　　要：以带有黄瓜绿斑驳花叶病毒的黄瓜叶片为材料,应用双抗体夹心酶联免疫吸附测定法(DAS-ELISA)、反转录-聚合酶链式反应(RT-PCR)、核酸斑点杂交(NASH)技术,进行检测灵敏度比较研究。3种方法检测灵敏度试验结果表明:DAS-ELISA、RT-PCR和斑点杂交可以检测出黄瓜病叶的最小量分别是39.6μg、0.25ng和5ng。RT-PCR检测灵敏度高于地高辛标记的核酸斑点杂交技术,DAS-ELISA检测灵敏度最低。Cucumber green mottle mosaic virus in virus-infected cucumber leaves was detected by DAS-ELISA、RT-PCR and NASH techniques and the sensitivities of detection were compared after the total RNA extracted from 0.1 g virus-infected leaves were diluted gradient 。The minimum weight of virus-infected cucumber leaves detected by DAS-ELISA, RT-PCR and NASH were 39.6 μg, 0.25 ng and 5 ng respectively. The results showed that the detection of RT-PCR was the most sensitive; NASH was more sensitive than DAS-ELISA. NASH technique can be replaced of RT-PCR used in metrical quarantine departments by comparison of sensitivities of three detect techniques.

关 键 词：黄瓜绿斑驳花叶病毒 反转录-聚合酶链式反应 双抗夹心酶联免疫吸附测定 核酸斑点杂交 检测技术 

分 类 号：S432.1[农业科学—植物病理学]