製作B型抗諧振反射光波導表面電漿子共振生物感測元件檢測登革熱病毒DNA之固定化與雜交

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。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。感測元件的所有設計與製程步驟會在論文中討論。生物感測實驗部分，進行登革熱病毒DNA固定化與雜交反應之即時檢測，為了有效增加固定化的效率，在此加入氯化鈉於DNA水溶液以及緩衝溶液中，而登革熱DNA分子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DNA分子DENV2-T之互補DNA序列與DENV2-P進行專一性鍵結。實驗結果顯示所設計與製作的元件可有效地應用此類檢測。

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-throughput and highly surface-sensitive platform to detect the biomolecular interactions. The waveguides in front and rear of the SPR sensing region have symmetric cladding structure to prevent from being affected by environmental changes, and sensing region is configured with a liquid flow channel which assists bioagents in attaching to the gold surface for reaction. The total design of our sensor chip and the fabrication process are discussed. For the bioassay experiments, the real time detection of dengue virus DNA immobilization and hybridization was studied. In order to enhance the efficiency of immobilization, NaCl is added to the DNA solution and the buffer solution. The dengue virus DNA probe DENV2-P was modified with a thiol group at one end to achieve immobilization on the Au surface, while the DNA target DENV2-T utilized the complementary sequence to be bound to the immobilized probe. The experimental results show that our devices can be effectively applied for this type of detection.