中文摘要：柑橘黄花叶病毒(CYMV)是花椰菜病毒科杆状DNA病毒属成员之一。该病毒是柑橘黄花叶病害的致病因子，可使柑橘减产。因为病毒是通过嫁接进行无性繁殖的，所以开发基于血清学技术的高通量诊断方法是生产健康脱毒种植材料的前提。作者研究介绍了可抵抗由CYMV ORF-II 编码的纯化重组病毒相关蛋白（rVAP）的兔源多克隆抗体的开发。本研究是第一个关于以重组病毒相关蛋白为融合蛋白制备多克隆抗血清的报道，这种多克隆抗血清可用于ELISA和IC-PCR法筛选感染CYMV的植株。这些免疫诊断方法可有效应用于柑橘常规检测和检疫。
外文摘要：Citrus yellow mosaic virus (CYMV) is a member of genus Badnavirus of the family Caulimoviridae. It is the causal agent of citrus yellow mosaic disease in citrus and causes reduction in yield. As the virus is vegetative propagated by grafting, development of high-throughput diagnosis methods based on serological techniques is a prerequisite for production of healthy virus-free planting material. The current study describes the development of polyclonal antibodies raised in rabbits against purified recombinant virion-associated protein (rVAP) encoded by ORF-II of CYMV. The specificity of developed antiserum was evaluated in immunosorbent electron microscopy (ISEM), antigen-coated plate-enzyme linked immunosorbent assay (ACP-ELISA) and immunocapture PCR (IC-PCR). The antiserum specifically reacted up to a dilution of 1: 2000 in ACP-ELISA for detection of CYMV-infected plants. The antiserum was validated by screening CYMV-infected plants maintained in the glass house through ACP-ELISA. To the best for our knowledge, this is the first report on production of polyclonal antiserum using recombinant virion-associated protein as fusion protein, which could be used for screening CYMV-infected plants by ELISA and IC-PCR. These immunodiagnostic methods can be effectively employed in routine indexing of citrus and in quarantine process.
外文关键词：Citrus yellow mosaic virus; Virion-associated protein; ACP-ELISA; IC-PCR
作者：Kumar, P. Vignesh; Sharma, Susheel Kumar; Bullet, Narayan Rishi; 等
作者单位：印度农业研究委员会
期刊名称：3 BIOTECH
期刊影响因子：1.361
出版年份：2017
出版刊次：12
点击下载：通过多克隆抗体与病毒相关蛋白重组表达进行柑橘黄花叶病毒有效免疫诊断的研究