中文摘要：

【目的】制备抗西瓜花叶病毒（Watermelon mosaic virus,WMV）的特异性单克隆抗体,并以其为核心建立能快速有效地检测WMV的血清学方法,从而为中国田间西瓜花叶病毒病的诊断和检测、预测预警及科学防控体系的建立提供物质和技术支撑。【方法】用提纯的WMV病毒粒子免疫BALB/c小鼠,经细胞融合和细胞培养、抗体筛选和细胞克隆等杂交瘤细胞技术,获得能稳定分泌抗WMV单克隆抗体的杂交瘤细胞株,将杂交瘤细胞注射入BALB/c小鼠腹腔制备其单抗腹水,并以制备的单抗为核心建立能准确、特异、灵敏地检测田间植物中WMV的ACP-ELISA、DAS-ELISA、dot-ELISA、Tissue blot-ELISA和IC-RT-PCR方法,以及能检测单头传毒介体蚜虫体内WMV的dot-ELISA方法。【结果】3株能稳定分泌WMV单克隆抗体的杂交瘤细胞株（2C8、15A8和16C12）及其单抗腹水被制备,3株杂交瘤细胞分泌的单抗腹水的间接ELISA效价均达到了10^-6以上,抗体类型及亚类均为IgG1、kappa轻链。Western blot分析发现,这3个单抗均与WMV的外壳蛋白亚基有特异性反应。灵敏度分析结果表明,ACP-ELISA、DAS-ELISA、dot-ELISA和IC-RT-PCR方法检测WMV病叶的灵敏度分别达到1﹕163 840、1﹕327 680、1﹕5 120和1﹕1 310 720倍稀释（w/v,g/mL）。特异性分析结果表明,以这3个单抗为核心建立的ACP-ELISA、DAS-ELISA、dot-ELISA、Tissue blot-ELISA和IC-RT-PCR 5种血清学检测方法均能与感染WMV的病叶发生特异性免疫反应,而与感染小西葫芦黄花叶病毒（Zucchini yellow mosaic virus,ZYMV）、黄瓜花叶病毒（Cucumber mosaic virus,CMV）、黄瓜绿斑驳花叶病毒（Cucumber green mottle mosaic virus,CGMMV）、马铃薯Y病毒（Potato virus Y,PVY）的植物样品、健康白南瓜、西瓜、葫芦和烟草的植物组织均呈阴性反应,且dot-ELISA方法还能特异性地检测单头蚜虫体内的WMV,而在检测无毒蚜虫时呈阴性反应。利用建立的血清学检测?

英文摘要：

【Objective】The aim of this study is to prepare monoclonal antibodies （Mabs） against Watermelon mosaic virus （WMV） and develop effective serological assays for rapid and reliable virus detection, and to provide technology and materiel for diagnosis and detection, forecast and early warning and establishment of a scientific prevention and control system of the WMV disease.【Method】Using the purified WMV particles as an immunogen, hybridoma lines secreting Mabs specific for WMV were obtained via cell fusion, cell culture, antibody detection and cell cloning. The hybridomas were injected intraperitoneally into BALB/c mice to produce Mab-containing ascitic fluids. Based on the prepared Mabs, five detection assays, ACP-ELISA, DAS-ELISA, dot-ELISA, Tissue blot-ELISA and IC-RT-PCR were developed for accurately, sensitively and specifically detecting WMV in field plant samples. Besides, a dot-ELISA for specifically detecting WMV in individual viruliferous aphid was established.【Result】Three hybridoma lines （2C8, 15A8 and 16C12） steadily secreting Mabs specific for WMV and their Mab-containing ascitic fluids were produced. The titers of ascitic fluids of Mabs were up to 10^-6 by indirect-ELISA. All these Mabs belong to IgG1 isotype, κ light chain. Western blot analyses indicated that all these three Mabs could specifically react with the coat protein of WMV. The ACP-ELISA, DAS-ELISA, dot-ELISA and IC-RT-PCR could detect WMV in infected plant crude extracts diluted up to 1﹕163 840, 1﹕327 680, 1﹕5 120 and 1﹕1 310 720 （w/v, g/mL）, respectively. And the specificity analyses demonstrated that the developed ACP-ELISA, DAS-ELISA, dot-ELISA, Tissue blot-ELISA and IC-RT-PCR had strongly positive immune reactions with WMV-infected plant tissues, but had negative reactions with healthy, ZYMV-, CMV-, CGMMV-infected cucurbitaceous plant tissues or PVY-infected tobacco plant tissues. Besides, the developed dot-ELISA for detecting vector sample had a strongly positive immune reaction with individual vi