| 于子翔1, 李丽2, 俞禄珍1, 杨翠云1, 吴建祥3, 黄卫昌2, 于翠1*.建兰花叶病毒和齿兰环斑病毒联合免疫胶体金试纸条的研制及应用[J].植物保护,2017,43(6):139-143. |
| 建兰花叶病毒和齿兰环斑病毒联合免疫胶体金试纸条的研制及应用 |
| Complex gold immunochromatography assay and application for rapid detection of Cymbidium mosaic virus and Odontoglossum ringspot virus |
| 投稿时间：2017-03-28 修订日期：2017-05-16 |
| DOI： |
| 中文关键词: 建兰花叶病毒 齿兰环斑病毒 联合免疫胶体金试纸条 |
| 英文关键词:Cymbidium mosaic virus Odontoglossum ringspot virus complex gold immunochromatography assay |
| 基金项目:上海市绿化和市容管理局(G152427) |
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| 中文摘要: |
| 为了实现兰花种植苗圃和口岸一线直接检测样品中建兰花叶病毒Cymbidium mosaic virus(CymMV)和齿兰环斑病毒Odontoglossum ringspot virus (ORSV)的目的,我们研制了上述两种病毒联合免疫胶体金检测试纸条。采用柠檬酸三钠还原法制备胶体金颗粒,标记单抗C10并固定于胶金垫上; 将抗体5B7和4F3分别固定于硝酸纤维素膜上作为检测线,将羊抗鼠抗体包被固定于硝酸纤维素膜上作为质控线,经优化后组装胶体金试纸条。结果表明研制的联合试纸条特异性强,能够在10 min之内同时检出两种病毒,操作简便。对两个种植苗圃的93份兰花样品进行实际检测所得结果与ELISA的检测结果符合性好(Kappa值分别为84.6%和86.7%),能满足兰花样品中建兰花叶病毒和齿兰环斑病毒的初筛要求。 |
| 英文摘要: |
| In order to directly detect Cymbidium mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV) in planting garden and import port, we developed gold-immunochromatographic assay stripe kit. Tri-sodium citrate with aqueous gold chloride was warmed up and mixed to make colloidal gold particles. Monoclonal antibody (McAb) C10 was selected to conjugate with colloidal gold as the detector antibody that was spotted on a conjugated pad, and the McAb 5B7 and 4F3 were used as capture antibody at the test line (T) respectively. Goat anti-mouse IgG antibody was used capture antibody at the control line (C). After an optimized process, the assay stripes were packaged. The result indicated that the viruses could be specifically detected on immune-strip within 10 minutes. 93 samples from Chenshan Botanical Garden and Sunqiao Agricultural Garden were tested, and the results were compared with those obtained by DAS-ELISA in order to examine the reliability. The two methods showed significant consistence (Kappa=84.6% and 86.7 % of CymMV and ORSV, respectively). The established gold-immunochromatographic assay could meet the requirements of primary screening of the two viruses in the gardens and the ports. |
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