以蠟燭為熱源進行核酸聚合酶連鎖反應

Abstract

目前商用的聚合酶連鎖反應（Polymerase chain reaction，PCR）儀器都需要電源驅動，在電力供給不穩定的第三世界國家很難進行PCR 的疾病檢測。本論文目的是用蠟燭代替熱源，在不同環境溫度下進行毛細管熱對流聚合酶連鎖反應（Capillary Convective Polymerase Chain Reaction，CCPCR）。此機制僅需使用單一溫控加熱玻璃毛細管底部，可成功進行DNA擴增，加熱系統部分使用蠟燭加熱鋁罐容器內的水，利用產生水蒸氣來維持玻璃毛細管底部溫度92℃。實驗結果顯示蠟燭CCPCR可成功於30分鐘擴增HBV 122-base pair DNA片段。靈敏度可達DNA濃度5 copies/tube。在7∼45℃環境溫度下，調整玻璃毛細管內的反應試劑體積，可成功進行DNA濃度5×105 copies/tube蠟燭CCPCR擴增。本研究提案不需要電源驅動的PCR疾病檢測技術，希望改善第三世界國家醫療發展。

Conventional polymerase chain reaction (PCR) instruments need power supply, so it is difficult to perform PCR for disease detection in the third world countries due to unreliable electricity supply. The purpose of this study is using candles as a substitute for a power supply to conduct the capillary convective polymerase chain reaction (CCPCR) at the different ambient temperatures. CCPCR mechanism only uses one single temperature controller, and heats the bottom of glass capillary tube in which the sample is contained. The heating system uses a candle light to heat the water in the aluminum container, and the bottom temperature of glass capillary tube is maintained at about 92℃ by the generated water vapor. The experimental results show the candle CCPCR is successful in the DNA amplification of HBV 122- base pair DNA fragment for 30 minutes. The detection sensitivity experiments of CCPCR can reach at an initial DNA concentration to 5 copies/tube. At an ambient temperature from 7℃ to 45 ℃,we are successful in the candle CCPCR amplification at the DNA concentration of 5×105 copies / tube by adjusting the reagent volume into the glass capillary tube. This study proposes the PCR for disease detection technology without the power supply, and we hope to improve the medical progress of the third world countries.