施加直流電之側向流免疫層析法對禽流感序列之快速偵測

Development of Lateral-Flow Immunochromatography Detection on the Gene Sequence of Avian Influenza by Applying A Direct Electric Field

陳志宏(Chih-Hung Chen)

指導教授 ： 吳瑞璋

中原大學/工學院/化學工程研究所/碩士(2013年)

https://doi.org/10.6840/cycu201300719

摘要

中文摘要
此研究示範了一種簡單且方便就可達到增強測向流薄膜試紙條
上對禽流感H5 DNA 的偵測訊號。在pH 8.5 及抗體濃度為25μg/ml
的條件下，奈米金顆粒與目標抗體具有最佳的結合效果，當H5 目標
DNA 被測試線上的DNA 探針所捕捉，此複合物則會產生可用肉眼判
讀的酒紅色偵測訊號。其原理是用一直流電場跨接於薄膜測向流試紙
條兩端，來誘導DNA 在膜上的電泳現象。隨著電壓的增強，所得的
偵測訊號也會跟著增強。以阿拉伯芥Pda13015、人類蛋白PSMA5 與
禽流感H5 基因在試紙條上做測試，所得的偵測結果顯示了此試紙條
對禽流感H5 DNA 具有很好的選擇性。此外，透過一系列不同待測
物濃度的偵測結果，也確認了有效的實驗再現性。在12V 下，所得
測向流薄膜試紙條的偵測極限可比無電壓的情況下增幅10 倍。由這
些結果來看，施加的直流電場可有效增加偵測訊號的強度。

關鍵字

快速偵測 ； 直流電 ； 側向流免疫層析法 ； 禽流感序列

並列摘要

Abstract
This study demonstrated a simple and convenient approach to enhance
the detection signals of the Avian-Influenza H5 DNA on the
membrane-based later-flow strips. The optimal antibody concentration
was determined as 25μg/ml at pH=8.5 for the conjugation between the
gold nano-particles and the target antibody. This conjugated complex
shows a visible red-wine color when H5 target DNA was captured by
DNA probe on the test line. A direct electric field was applied over two
ends of the strips to induce an electrophoresis of DNA through the strips.
By increasing the voltage, signal intensity was also enhanced. The
experimental results showed that the strips had an excellent selectivity of
Avian Influenza H5 from other two gene species of Arabidopsis thaliana
Pda13015 and human protein PSMA5. Effective reproducibility of
experiments was also verified over various analyte concentrations to
confirm the result validity. The application of 12 volts on the
membrane-based later-flow strip indicated that the detection limit could
be enhanced in 10 folds of the no-voltage strip. The results indicated that
a direct electric field offered a promising signal enhancement of detection
readings.

並列關鍵字

Avian Influenza ； Lateral-Flow Immunochromatography ； Direct Electric Field

參考文獻

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