2005-01-012024-05-15https://scholars.lib.ntu.edu.tw/handle/123456789/663711摘要：植物病毒病害是目前台灣所有植物病害中最難以防治的病害之一,除了病害防治為 最終目的外,早期正確而快速簡易的病毒病診斷,在病害管理中扮演著非常重要且決 定性的角色.尤其未來加入WTO後,農產品的進出口檢疫將是一必要的工作,而病害 之快速診斷法更是不可或缺.近年分子生物技術與電子技術結合,發展出生物晶片 （Bio-chip）檢測法,在生物醫學上已漸被開發利用.生物晶片在植物病毒診斷技術 之建立,在未來加入WTO後,對於大量之檢驗樣品,將可以達到快速、大量、精確之 目的.病毒之核酸探針之製備,核酸探針之標定,核酸探針之點陣,核酸雜合,生物晶片 法之效果評估.預期效益在於完成以生物晶片診斷植物病毒技術之建立,完成以生物 晶片和其他診斷植物病毒技術之比較,提供以生物晶片診斷技術作為大量檢疫可行 性評估.Potyvirus為植物病毒之中最大且具經濟重要性的一群，為研發能鑑定各種 potyvirus之植物病毒快速鑑定晶片，並將研究成果轉移給國內防疫與檢疫單位使 用，本計畫今年將繼續由GenBank收集並分析重要potyvirus的序列資料，設計並製 備能鑑定其他potyvir<br> Abstract: Plant virus diseases are most difficult problems to control. Besides control methods, simple and fast diagnosis of disease in the early stage is most important in the disease management. Especially after jointing into WTO, quarantine for import agricultural products is essential. Therefore fast diagnosis systems have to be developed. Biochip has been used in the biomedical susseciqully. In the future biochip will be perfectly applied in the diagnosis of plant viruses. The project includes preparation of cDNA probes, labeling of cDNA hybridization and access of biochip.Potyvirus is the biggest and the most economically important group of plant viruses. In order to develop a rapid identification chip for potyviruses and provide the chip to the authorities of plant inspection and quarantine, the following works will be executed this year. (1) Continuously collect and analyze the sequences of potyviruses, design and prepare the oligo probes for other potyviruses. (2) According to the sequence of cDNA clones and the multiple alignment results of potyviruses, design and prepare the cDNA probes. (3) Study the specificity and sensitivity of the probes to the cDNA clones of potyviruses by reverse dot blot hybridization. Establish the best detection procedure and shorten the required time. (4) Check the detection accuracy of the rapid identification chip on potyvirus-infecting plant materials. By the end of the year, there will be another 15 kinds of probes designed, and the specificity and sensitivity of individual probes will be studied. Plant disease control and quarantine on Fusarium pathogens, resulted from the free world trade in the agricultural products, will undoubtedly be more complicated and difficult. The identification of Fusarium phytopathogenic species has been based mainly on the morphological characteristics and has been considered very troublesome. Development of a chip-based detection is an attractive alternative to cope with the problems to come. This project has aimed to examin the RAPD and AFLP molecular polymorphisms, and to develop formal speciales-specific nucleic acid probes to provide a rapid molecular detection and diagnosis. We expect to obtain more sets of DNA probes specific to different formal speciales of Fusarium causing crop wilts in Taiwan in 2004. Deployment of a DNA microarray-based pathogen detection and disease diagnosis system is very expensive. Therefore, we intend to develop a processing chip that can electrophoretically separate and electrochemically detect the DNA fragments. In 2004, we expect to develop and a μ-CE/ECD chip capable of performing sizing and detection of the above-mentioned formal speciales-specific RAPD- and AFLPproducts. The goal of this research project is to develop a rapid detection technique for important fungal pathogens- Fusarium spp. This rapid detection technique is based on the designed specific PCR fragments of the specific pathogen as the target DNAs,基因晶片豬病毒疾病馬鈴薯Y屬病毒鐮孢菌μCE/ECD晶片聚合&#37238連鎖反應大腸桿菌生物晶片DNA晶片Gene chipSwine viral diseasespotyvirusFusarium spp.μCE/ECD chipPolymerase chain reactionE. coli biochipDNA chip