DNA萃取和即時聚合酶鏈鎖反應於數位微流體晶片

Abstract

本文是利用數位微流體 (Digital microfluidic，DMF) 技術整合DNA萃取和即時聚合酶鏈鎖反應 (Real-time polymerase chain reaction，qPCR) 於晶片內。為了使point-of-care更加普及化，未來必須發展出一個個人裝置在不需要臨床人員下，能夠在任何時候和任何地方都可以即時檢測疾病，隨時都能掌握自己的健康狀況，最關鍵的技術就在DNA萃取和qPCR。在此晶片上可利用交流電產生介電濕潤 (Electrowetting on dielectric，EWOD) 來移動人體血液跟生物試劑套件混合來萃取血液裡的DNA，在100 nL的血液裡萃取出4.45 ng/μL濃度的DNA。跟傳統的DNA萃取比較，萃取時間減少75 %的反應時間，從120分鐘縮短至30分鐘，試劑消耗減少99.95 %的體積，從4222

This thesis reports the implementation of DNA extraction and real-time PCR (qPCR) on a digital microfluidic (DMF) device. We aim to develop a personalized point-of-care device for molecular diagnosis from human whole blood and commercial reagents kits driven by electrowetting-on-dielectric (EWOD) on a DMF device for DNA extraction and qPCR. The results from on-chip DNA extraction protocols were validated and quantified. In comparison wiht the traditional DNA extraction procedures in tubes, our on-chip extraction starting from a 100 nL whole blood obtained 52.8 % DNA concenttion at 4.45 ng/µL, required 25 % reaction time (from 120 min to 30 min), consumed 0.05 % volume of the entire reagents (from 4222