本研究提出新型DNA雜交反應訊號分析方法，結合微機電技術與生物晶片科技，同時利用奈米金-鏈黴卵白素複合體(gold-streptavidin)與銀析出增強溶液放大反應偵測訊號，並配合金屬銀析出沉澱之反應，以光學式與阻抗式偵測系統來量測訊號，有別於傳統以酵素、螢光或放射性同位素標識呈色方法，建立一新型DNA雜交反應分析系統，提供另一種有效且方便的DNA分析檢測方式。在DNA樣品選擇上，選擇以鮑氏不動桿菌(Acinetobacter baumannii)作為DNA樣品來源，主要是因為臨床上鮑氏不動桿菌發生院內感染率很高。

此外，在微晶片上執行DNA雜交反應具有改良多樣性、低樣品用量與高靈敏度等特性，且可以縮短偵測反應的時間。實驗架構上，同時利用生物素-鏈黴卵白素(biotin-streptavidin)具有專一性鍵結能力，且奈米粒子具有催化銀析出增強溶液特性，作為放大訊號理論基礎，並採用3-環氧丙三甲基矽氧烷(3-glycidoxypropyl-trimethoxysilane, 3-GPS)進行晶片表面化學修飾，並將具有專一性序列探針固定在晶片表面，對待測DNA樣品作定性與定量之分析，加入銀析出增強溶液，來增顯雜交反應的偵測訊號，同時利用光學式灰階掃描與阻抗式電阻量測二種方法，來擷取實驗結果的影像及數據。

本研究方法已可完成DNA雜交的步驟與目標，光學式與阻抗式對DNA樣品之偵測靈敏度分別達到8.25 ng/mL與8.25×10-1 ng/mL，且使用的偵測方式也可搭配銀析出現象產生可供檢測之訊號，大幅縮短檢驗時間。此外，在實驗中將討論DNA雜交參數與靈敏度等問題，同時利用生物分子特性、化學性修飾、奈米粒子技術與銀訊號增強等方法，建立一個全新、快速且靈敏之DNA雜交訊號分析系統，未來將提供DNA分析檢測一個全新的方向與思維。

This study reports a novel DNA hybridization assay method which binding micro-electro mechanical systems (MEMS) and biochip technology. Using gold-streptavidin and silver enhancement solution, as a catalyst for silver precipitation, magnify hybridization detection signals. Finally, using optical and electric system detect signals. Compared with traditional labeling methods such as enzyme, fluorescence or radioisotope, we supply a new DNA hybridization assay system that is simpler, cheaper and more convenient. We choose Acinetobacter baumannii as DNA sample source, because the occurrence of bacteremia by Acinetobacter baumannii for glucose nonfermenters on clinical diagnosis is maximum.

Furthermore, DNA hybridization reacted on micro-chip having advantage of low sample waste, high sensitivity and short detection time. In experiment scheme, we combine the specificity binding ability of biotin-streptavidin and catalyst for silver precipitation ability of nanoparticles as the foundation theory. At the same time, used 3-glycidoxypropyl-trimethoxysilane (3-GPS) to modify the surface of slide chip and immobilize DNA probes with specificity sequence on slide chip. DNA sample are used to quantify and qualify test rely on silver enhancement method with optical and electric system to get images and data of experiment results.

The relationship between sample concentration and detection signal is discussed and the detection limit for the DNA sample are 8.25 ng/mL and 8.25×10-1 ng/mL in optical and electric system respectively. In addition, the parameter, specificity and sensitivity of the DNA hybridization assay chip are discussed. We combine bio-molecule, chemical modification, nano-technology and silver enhancement to achieve a novel DNA hybridization system. The high applicability and biochemical efficiency of this study can provide an alternative for rapid, sensitive and convenient DNA hybridization assay method.

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