核酸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 of Avian Influenza

楊環菁(Huang-Chin Yang)

指導教授 ： 吳瑞璋

中原大學/工學院/化學工程學系/碩士(2008年)

https://doi.org/10.6840/cycu200800425

摘要

免疫層析試紙分析法（immunochromatography）是現代快速生物感測的重要方法之一，其原理是以抗體、抗原間專一性與免疫親和力為基礎，而在此實驗中則利用DNA-ligand 和anti-ligand antibody 替代抗體、抗原分析法以偵測禽流感DNA 的序列，並利用雜合反應做為檢測基礎，其檢測方式非常簡便。目標物DNA序列首先以聚合酶鏈鎖反應（PCR）擴增且標誌上特殊標定物，使待測樣品在硝化纖維膜流動時，藉由被anti-ligand antibody捕捉，經由膠體金得顯色可判讀出是否含有禽流感之特殊序列。本論文也發現在薄膜兩端加入直流電壓可增進信號強度。最後得到最佳條件下的偵測極限（detection limit）為100pg。在本論文中也詳細討論到在條件變化下，偵測極限會隨之變化，並指出所遇到之問題及其解決方式。

關鍵字

DNA檢測 ； 側向流免疫層析 ； 免疫分析 ； 禽流感 ； 硝化纖維膜

並列摘要

The later-flow immunochromatography is one of modern rapid detection methodologies using the principle of specific immunity between antigens and their affinity antibodies. Instead of antigen-antibody immunoassay, this study employed the binding specificity between DNA-ligand and anti-ligand body to detect Avian Influenza by the hybridization reaction in a rapid and convenient manner. The target DNA amplified by the Polymerase Chain Reaction (PCR) and labeled with a specific ligand was loaded onto nitrocellulose membrane to laterally flow through and be captured by an anti-ligand antibody. The detection result was designed to be visually read out by gold-sol to identify distinct sorts of Avian Influenza. A direct voltage applied over the membrane was verified effective to enhance the detection limit. The detection limit was determined as about 100pg but was change at different conditions.

並列關鍵字

DNA detection ； Avian Influenza ； lateral-flow Immunochromatography ； immunoassay ； nitrocellulose membrane

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