利用表面電漿子共振矽基片B型抗諧振反射光波導生物感測元件於登革熱病毒DNA之雜交與專一性之檢測

Abstract

本論文研究以B型抗諧振反射光波導(ARROW-B)表面電漿子共振(SPR)生物感測器量測登革熱病毒DNA之雜交及專一性。和以往利用螢光標定於DNA探針之偵測不同，本生物感測器有免標定、即時偵測、低成本與步驟簡單快速之優點。利用互補序列的探針及樣品做DNA雜交的分析對於醫學診斷是很重要的技術。但是傳統分析技術過於複雜並且耗時，因此希望利用本元件提供一個檢測DNA上更好的平台。本元件利用表面電漿子共振現象作為生物感測的方式，藉由流道系統幫助生物分子附著於感測區表面進行反應,當感測區域中表面等效折射率的改變時，會造成通過波導的光強度訊號有所變化。登革熱單股DNA分子DENV-P利用末端修飾的硫醇基先行固定於金表面之後，再利用登革熱單股DNA分子DENV-T之互補DNA序列與DENV-P進行專一性鍵結。此外，為了驗證其專一性,也利用Oligos進行非特異性鍵結之測試。實驗結果顯示此生物感測元件能有效地即時檢測此DNA的雜交及專一性反應。

In this study, we detected dengue virus DNA hybridization and specificity by using antiresonant reflecting optical waveguide B (ARROW-B) surface plasmon resonance (SPR) biosensors. In contrast to traditional fluorescence techniques, our sensor has many advantages including label-free, real-time, low cost and rapid process. DNA hybridization analysis, based on the complementary base pairing of target and probe DNAs, has become increasingly important in diagnostic applications, but traditional techniques are complicated and spend more time. For this reason, we hope to provide a better platform to detection with ARROW-B SPR sensors. This sensor was configured with a liquid flow channel which assists bioagents in attaching to the gold surface for reaction. When we injected different reagents into the chamber, it caused the changes of effective index, giving rise to variations of light power. The dengue virus DNA probe (DENV-P) was modified with a thiol group at one end to achieve effective immobilization on the Au surface, while the dengue virus DNA target (DENV-T) utilized the complementary sequence to bind to the immobilized probe. Besides, we utilized Oligos to conduct non-specific binding test to verify the specificity of DNA hybridization. In summary, the measurement results have shown that ARROW-B SPR biosensors could be applied to detect the dengue virus DNA hybridization and specificity in real time.