本研究開發一自動化微流體晶片系統，藉由微型磁珠上對登革熱病毒極高專一性的抗體可以針對登革熱病毒或是因感染登革熱所產生的免疫球蛋白M (Immunoglobulin M, IgM)和免疫球蛋白G (Immunoglobulin G, IgG)作結合，進行以磁珠為基礎的免疫分析法檢測流程，可同時偵測登革熱病毒專一的IgM和IgG。相較於傳統檢測方法，酵素連結免疫分析法 (Enzyme-linked immunosorbent assay, ELISA)是用平面式的反應槽進行檢體抗原抗體的結合反應，有著繁瑣的人為操作程序，耗費相當多的反應時間。將微型磁珠應用檢體之萃取，並在本晶片上自動地完成抗原抗體的接合反應以及檢體的螢光偵測，可以減少人為操作的不定性，更可大幅縮短操作時間與步驟，其具有高檢測靈敏度、可拋棄式、可攜帶性、低樣品及檢體消耗量、低耗能、體積小以及成本低等優點，相較傳統分析檢測技術下，有著突破性的發展價值。
此微流體晶片系統中包含微型幫浦、薄膜式微混合器以及微型線圈等元件。其中薄膜式微混合器操作條件為1.37 Hz與通入之壓縮氣壓其壓為20 psi時，可於6秒內達到約莫96 %的混合效果，可使檢體於最短的時間內有最佳的混合效率並且有效的提升檢測的靈敏度。而所有元件將整合於一微型晶片系統上，共包含一薄膜式檢體混合模組、二雙向式微流體傳輸模組、一微陣列式環型磁線圈以及三個單向微流體傳輸模組和兩個螢光偵測區，以達到自動化傳輸流體、混合的功能，並於最後將微流體系統晶片應用於檢體內IgG和IgM檢測。
本研究成功地使偵測極限由傳統酵素連結免疫分析法的0.8 ng，提高到21 pg，反應時間也由4小時縮短至30分，而使用螢光系統取代傳統酵素連結免疫分析法的酵素呈色可以提升整個系統的靈敏度。另外，實驗結果也顯示利用磁珠為基礎的免疫分析法確實可以偵測流感病毒，使靈敏度從傳統的1.28 HAU增加到1.28×10-5HAU。本研究也成功的實驗證明了利用免疫檢測微流體晶片系統在生醫檢測分析上確實大大地提升檢測的效能與靈敏度。

This study reports an integrated microfluidic system which utilizes virus-bound magnetic-bead complexes for rapid serological analysis of antibodies associated with an infection by the dengue virus. This new microfluidic system integrates one-way micropumps, a four-membrane-type micromixer, two-way micropumps and an on-chip microcoil array in order to simultaneously perform the rapid detection of immunoglobulin G (IgG) and immunoglobulin M (IgM). An IgM/IgG titer in serum is used to confirm the presence of dengue virus infection. By utilizing microfluidic technologies and virus-bound magnetic beads, IgG and IgM in the serum samples are first captured. This is followed by purification and isolation of these beads utilizing a magnetic field generated from the on-chip microcoil array. Any interfering substances in the biological fluids are washed away automatically by the flow generated by the integrated pneumatic pumps. The fluorescence-labelled secondary antibodies are then bound onto the surface of the IgG/IgM complex attached onto the magnetic beads. Finally, the entire magnetic complex sandwich is transported automatically into a sample detection chamber. The optical signals are then measured and analyzed by an optical detection module. The entire process is performed automatically on a single chip within 30 minutes, which is only 1/8th of the time required for a traditional method. More importantly, the detection limit has been improved to 21 pg, which is about 38 times better when compared to traditional methods. In addition, the magnetic-bead-based immunoassay also can detect influenza virus, the sensitivity has been enhanced to 1.28×10-5 HAU. This integrated system may provide a powerful platform for the rapid diagnosis of dengue virus infection and other types of infectious diseases.

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