利用B型抗諧振反射光波導表面電漿子共振生物感測元件進行登革熱病毒核酸序列雜交之檢測

Abstract

本論文研究適用於水中環境的B型抗諧振反射光波導(ARROW-B)表面電漿子共振(SPR)生化感測元件。由於此生物感測器具有免標定、高效率以及對於金表面所固定的生物分子層之折射率變化具有高靈敏度等特性，因此能夠應用在即時感測表面生物分子之交互作用。此生物感測元件的設計，使用模擬分析光場在元件中的傳播特性，使其在水環境下具有最佳化的感測靈敏度。在感測區前後的光場傳播區設計具上下對稱的光波導隔離層，使元件能穩定導光而不受外界環境影響，並於感測區附加液體流道設計，幫助生物分子附著於感測區表面進行反應。生物感測實驗部分，進行登革熱病毒之專一性核酸序列雜交反應之即時檢測，登革熱病毒檢測之探針序列單股DNA(DENV2-P)利用末端修飾的硫醇基先行固定於金表面之後，再利用登革熱病毒之專一性分子DENV2-T與DENV2-P進行專一性雜交實驗結果顯示，此生物感測元件能有效地即時檢測此DNA雜交反應。

In this study, an antiresonant reflecting optical waveguide of type B (ARROW-B) surface plasmon resonance (SPR) biosensor operating in the aqueous environment with a circulating-flow system has been investigated. The ARROW-B SPR biosensor is proposed to provide a label-free, high-efficiency and highly surface-sensitive platform to detect the bimolecular interactions. Modal characteristics of ARROW-B are analyzed with simulation and designed for obtaining optimum sensitivity in aqueous environment. The waveguides in the front and rear of the SPR sensing regions have symmetric cladding structure to prevent from being affected by environmental changes, and the sensing region is configured with a liquid flow channel which assists bioagents in attaching to the gold surface for reaction. For the bioassay experiments, the real time detection of specific nucleotide sequences hybridization of dengue virus was studied. The dengue virus DNA probe was modified with a thiol group at one end to achieve effective immobilization on the Au surface, while the DNA target utilized the complementary sequences to hybridize to the immobilized probe. In summary, the measurement results have shown that the ARROW-B SPR biosensors could be applied to detect the dengue virus ssDNA hybridization in real time.