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標題: 開發具產業應用潛力之分子檢測商業化服務系統平台-應用高輸出量(High Throughput)技術於重要農業害蟲分子檢測系統之開發
Development of High Throughput Molecular Detecting Systme for Rapid Identification of Improtant Insect Pests
作者: 路光暉
關鍵字: 生物技術;應用研究
前期計畫我們已針對頸吻蟬類(飛蝨、葉蟬和沫蟬等)、木蝨類、蚜蟲類和果實蠅類等四類重要農業害蟲,各挑選5~10 種我國植物防疫與檢疫工作上較具重要性的國內外蟲種,分別進行形態特徵與DNA 分子分析。其中形態部分已建立各蟲種之分類特徵,用以確認物種之界定;DNA 分析的部分,則以分析粒線體細胞色素I(cytochrome oxidase I, COI)基因區段及第二核醣間區(intergenicspacer 2, ITS2)等兩類DNA DNA 分子之變異,研發得一套具區辨害蟲種類功效之生物晶片(bio-chip)系統,並建立其檢測方法與標準操作流程。本期計畫將著重於加強此初步研發成果,包括(1)繼續自不同區位,如第一核醣間區(intergenic spacer 1, ITS1)或轉譯延展因子-1脈(elongation factor-1-alpha, EF1脈)等等,篩選探針序列,增加DNA 標誌的代表性,強化種類鑑定的專一性;(2)繼續測試如檢體DNA 的萃取方法、PCR 增幅的條件、探針序列位置、濃度與長度、雜合時間與溫度等等,進一步改良此晶片系統,建立更有效、穩定與快速的標準檢驗流程。由於本計畫研究成果將具獨創性與專利性,因此將具商業價值,本期計畫期間亦將尋求適當法人機構或私人產業作技術轉移與授權,進行害蟲種類的有償鑑定服務。

In previous study, we focused on four groups of insect pests, includingAuchenorryncha (e.g. planthoppers, leafhoppers, and froghoppers, etc.), psyllids,aphids, and tephritids, which are notable pests in agriculture. Five to ten species ofeach group were selected to approach the subjects using both morphological andmolecular analyses. Morphological characteristics were used for confirmation ofeach species for further molecular analysis. Based on the DNA sequence variation ofcytochrome oxidase (COI) and intergenic spacer 2 (ITS2) between species, we havedeveloped biochips and standard procedures for rapid identification of these fourgroups of pest insects. In this proposal, we propose a further improvement of ourprevious achievement mainly in two objectives: (1) to select different DNA regionssuch as intergenic spacer 1 (ITS1) and/or elongation factor-1脈(EF1脈) to screen morespecies-specific probes in order to increase accuracy of identification; (2) to furtherimprove the performance of our biochip systems by optimizing a variety of reactionparameters, including the methods of DNA extraction, the size, concentration, anddistribution of probes on the chip, the time and temperature for hybridization, etc.The achievement of current research will have its uniqueness and commercial valuewhich should be suitable for applying patent and the technology could be transferredto private institute.
其他識別: NSC97-2317-B005-009
Appears in Collections:昆蟲學系

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