2005-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/710432摘要：海芋為天南星科、馬蹄蓮屬多年生球根花卉，近幾年來深受消費者喜愛，栽培面積持續增加，是國內極具發展潛力之球根花卉。海芋栽培的限制因子之一為病毒性病害，病毒對於海芋的生長與觀賞品質都有不利的影響，且田間常發現有複合感染的情形，因此急需建立海芋病毒的快速檢測方法，以協助健康海芋種苗之生產。近年來台灣學者發現多種感染海芋的新病毒，研究成果領先世界各國，因此有關海芋病毒的研究極具市場價值和世界競爭性。我們將利用免疫層析檢測法的原理，以廣效性單元抗體配合專一性多元抗體或單元抗體，研發操作簡便、可同時偵測多種海芋病毒的快速診斷套組，這種新穎的病毒檢測觀念是我們首創的。本計畫預定以三年五個月的時間研發出可同時檢測5種國內常見的海芋potyvirus的快速診斷套組。為達成此目的將進行下列研究項目：(1)選殖和表現5種海芋病毒CLLV、DsMV、TuMV、ZaMV和ZaMMV的鞘蛋白基因；(2)分別製備和分析5種海芋病毒的專一性多元抗體；(3)製備和篩選5種海芋病毒的廣效性單元抗體；(4)純化病毒顆粒供篩選抗體和檢測使用；(5)分別製備和篩選5種海芋病毒的專一性單元抗體；(6)測試膠金標定單元抗體的條件；<br> Abstract: Calla lily (Zantedeschia spp.), belonging to the family Araceae, is a perennial bulbous flower and has become popular flower crops in Taiwan. One of the limiting factors for calla lily production is viral disease. When infected by viruses, the growth and flowering of calla lilies will be severely affected. Besides, mixed infections are frequently found in the field. Therefore, it is essential to establish rapid detection methods for calla viruses in order to produce virus-free seedlings. In the past few years, the research results of calla viral diseases in Taiwan are leading the world because several new viruses were identified by local scientists. For that reason, the project about calla lily virus will be valuable and important. We are going to utilize the concept of immunochromatographic assay, and use the genus-specific monoclonal antibody, the species-specific polyclonal antibody or monoclonal antibody to develop use-friendly rapid diagnosis kit for simultaneously detecting several calla lily viruses. This novel concept for multiple virus detection is first proposed by us. In order to achieve the goal of developing a rapid diagnosis kit for five calla lily-infecting potyviruses, we will perform the following research items: (1) clone and express the coat proteins of CLLV, DsMV, TuMV, ZaMV and ZaMMV; (2) separately prepare the polyclonal antibodies of CLLV, DsMV, TuMV, ZaMV and ZaMMV and analyze their specificity; (3) prepare and screen the genus-specific monoclonal antibody of CLLV, DsMV, TuMV, ZaMV and ZaMMV; (4) purify virus particles for antibody screening and detection test; (5) prepare and screen the species-specific monoclonal antibody of CLLV, DsMV, TuMV, ZaMV and ZaMMV; (6) test the condition of colloidal gold labeling for monoclonal antibody; (7) develop a convenient grinding tool and extraction solution for calla lily; and (8) finally develop a rapid diagnosis kit for calla lily viruses.海芋病毒檢測快速診斷套組Calla lilyVirusDetectionRapid diagnosis kit