|
|||||
|
|
| 番茄褐色皱果病毒胶体金免疫试纸条的研制 | |
| 引用本文： | 闫志勇,牟修岐,赵梅胜,马华玉,耿超,田延平,杨武杰,李向东.番茄褐色皱果病毒胶体金免疫试纸条的研制[J].植物病理学报,2022,52(6):976-983. |
| 作者姓名： | 闫志勇 牟修岐 赵梅胜 马华玉 耿超 田延平 杨武杰 李向东 |
| 作者单位： | 山东省农业微生物重点实验室,山东农业大学植物保护学院,泰安 271018; 山东省农业技术推广中心,济南 250014 |
| 基金项目： | 国家自然科学基金(32072387); 泰山学者建设工程(TS2017120023) |
| 摘 要： | 番茄褐色皱果病毒(tomato brown rugose fruit virus, ToBRFV)是一种新发病毒,严重威胁番茄的安全生产。为了快速、简便地检测该病毒,我们制备了ToBRFV胶体金免疫试纸条。本研究以ToBRFV粒子为免疫原,通过杂交瘤技术制备了17个抗ToBRFV的单克隆抗体。将不同单抗两两组合分别作为胶体金标记抗体和硝酸纤维素膜检测线上的捕获抗体,共获得272个配对组合的胶体金试纸条。通过特异性测定筛选到一组配对抗体制备的试纸条能够在5 min内特异识别ToBRFV,而与番茄斑驳花叶病毒、番茄花叶病毒、烟草花叶病毒、黄瓜花叶病毒、辣椒轻斑驳病毒、马铃薯X病毒、马铃薯Y病毒、番茄褪绿病毒、番茄斑萎病毒、番茄黄化曲叶病毒等无交叉反应。灵敏度分析表明,该试纸条可从稀释12 800倍的番茄叶片病汁液中检测到ToBRFV,也可检测到50 ng ToBRFV粒子。本研究制备的胶体金试纸条使用方便,灵敏度高,特异性强,适合田间大批量样品检测,可用于ToBRFV的精准监测及早期预警。 |
| 关 键 词： | 胶体金免疫试纸条 检测 特异性 番茄褐色皱果病毒 |
| 收稿时间： | 2021-09-21 |
Development of colloidal gold immunochromatographic strip for detection of tomato brown rugose fruit virus |
|
| YAN Zhiyong,MU Xiuqi,ZHAO Meisheng,MA Huayu,GENG Chao,TIAN Yanping,YANG Wujie,LI Xiangdong.Development of colloidal gold immunochromatographic strip for detection of tomato brown rugose fruit virus[J].Acta Phytopathologica Sinica,2022,52(6):976-983. | |
| Authors: | YAN Zhiyong MU Xiuqi ZHAO Meisheng MA Huayu GENG Chao TIAN Yanping YANG Wujie LI Xiangdong |
| Institution: | Shandong Provincial Key Laboratory of Agricultural Microbiology, College of Plant Protection, Shandong Agricultural University, Tai’an 271018, China; Shandong Provincial Agricultural Technology Extension Center, Ji’nan 250014, China |
| Abstract: | Tomato brown rugose fruit virus (ToBRFV) is an emerging virus and seriously threatens the safety of tomato production. To achieve rapid and simple detection of the virus, we developed a colloidal gold immunochromatographic strip for ToBRFV. Here, we used ToBRFV particles as the immunogen and produced 17 monoclonal antibody lines by hybridoma technology. Two different monoclonal antibodies were individually used for the colloidal gold-labeled antibody and the capture antibody on the detection line of the nitrocellulose membrane and combined to prepare colloidal gold immunochromatographic strips with 272 paired combinations. The immunochromatographic strip prepared with one paired antibodies could specifically recognize ToBRFV within 5 minutes, without reacting with tomato mottle mosaic virus, tomato mosaic virus, tobacco mosaic virus, cucumber mosaic virus, pepper mild mottle virus, potato virus X, potato virus Y,PVY, tomato chlorosis virus, tomato spotted wilt virus, or tomato yellow leaf curl virus. The sensitivity test results showed that the minimum detection threshold was 12 800 times for ToBRFV-infected tomato leaf extracts and 50 ng ToBRFV particles. The resultant colloidal gold immunochromatographic strip, which is sensitive and specific for ToBRFV, easy to use, and competent for large-scale sample detection, can be applied in the precise monitoring and early warning of ToBRFV. |
| Keywords: | colloidal gold immunochromatographic strip detection specificity tomato brown rugose fruit virus |
| 点击此处可从《植物病理学报》浏览原始摘要信息 | |
| 点击此处可从《植物病理学报》下载免费的PDF全文 | |