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標題: | 關於增強靈敏度的表面電漿子共振適配子感測器於檢測砷之研發 Development of a Sensitivity-Enhanced Surface Plasmon Resonance Aptasensor for Detection of Arsenic |
作者: | Letao Fan 范樂陶 |
指導教授: | 林啟萬(Chii-Wann Lin) |
關鍵字: | 表面電漿子共振,適配子,砷,DNA探針, surface plasmon resonance,aptamer,arsenic,DNA probe, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 由於地質礦物的自然分佈和工業污染,砷在世界範圍內一直是水質和土壤的主要污染源之一,嚴重危害著人類的生產和生活。就現有的檢測技術而言,大多數對砷的檢測都需要在實驗室內,並且利用複雜、精密的設備進行,儘管這些檢測方法都具備極低的最低檢測極限(LOD),但是對於臨場檢測和實時監控的應用,都非常的不切實際。因此,在主要針對農業用水、飲用水儲備水源和工業廢水監測的應用上,就需要開發一種具備高靈敏度,且最低檢測極限能夠滿足大多數需求的快速篩檢機制。同時,SELEX技術的發明,實現了針對特定標的物具有親和性的DNA適配子的開發與製造。基於M. Kim團隊1研發的對砷具有特定親和性的DNA適配子,本研究團隊得以進行針對砷的表面電漿子共振適配子感測器的開發。然而,我們在研究中發現,當適配子被固定在奈米金膜的表面時,其自身的二級結構會抑制其對檢測物的靈敏度。我們繼而開發了一個解決方法,即利用一段與適配子無功能區段互補的DNA序列與之結合,從而將適配子二級結構進行矯正,使其能夠更加充分的與檢測物反應。經過實驗證明,這個方法可以有效的提升系統靈敏度,降低最低檢測極限,並且不會對適配子的特異性產生影響。這個方法同樣在對臨場水質樣本的測試中證明有效,並且具備等同於標準品測試的再現性。本研究亦為利用互補DNA序列進行適配子結構與功能的調整提供了可能性,並且在未來藉由與實驗室先前針對其他水質重金屬污染物而開發的檢測技術相整合,亦可以進行多功能的晶片實驗室(lab-on-a-chip)之開發。 Arsenic has been contaminating water and soil worldwide due to mineral distribution and industrial pollution. The current detecting technologies are mostly lab-based with complex and delicate equipment, though with extremely low limit of detection (LOD), they are impractical for on-site screening and real-time monitoring. Thus a rapid screening method is in demand with high sensitivity and a LOD low enough for inspections of agricultural water, reservoir water and wastewater from industrial discharge. The introduction of Systematic Evolution of Ligands by Exponential Enrichment (SELEX) made it possible to develop DNA aptamers affinitive to assigned targets including arsenic. Based on the arsenic-affinitive aptamer developed by M. Kim’s research team1, we were able to develop a surface plasmon resonance aptasensor for detection of arsenic. However, during our research we discovered that the secondary structure of the DNA aptamer inhabited its own sensitivity to the target when being immobilized on the gold nano-film. So we developed a solution to bind a complementary DNA segment to the aptamer on its non-functional site to erect its secondary structure, allowing better exposure to the analytes, which has been proven effective with enhanced sensitivity, lowered LOD and no influence on specificity to the target. This method was also tested with field sample, resulting in decent reproducibility in performance. Our research also implies the viability in aptamer modification by complementary DNA segments and, along with previous researches on detection of other aqueous heavy metal ions by our lab, we are able to make another step forward to the development of an integrated multi-function lab-on-a-chip in the future. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59478 |
DOI: | 10.6342/NTU201700958 |
全文授權: | 有償授權 |
顯示於系所單位: | 醫學工程學研究所 |
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