開發側向流免疫分析試紙進行基質金屬蛋白酶偵測

Abstract

在 2010 年全球有 10.79% 的人口受牙周病的影響，也就是將近 7.5 億的人口總數，並造成約 539 億元的全球生產總值下降，造成巨大的經濟負擔。因此，為了解決牙周病衍生的各種社會問題，偵測早期牙周病並及時處理與治療，可大幅降低牙周病後期所需要的醫療開銷，降低國家在口腔疾病治療上的負擔。因此為了預防牙周病的發生，本論文開發了側向流免疫分析試紙，並以此裝置偵測牙周病的標準生物細胞因子-基質金屬蛋白酶 (Matrix metallopeptidase 9)。我們成功達成 (1) 金奈米粒子與 MMP-9 檢測抗體的鍵結；(2)MMP-9 抓捕抗體最佳化；(3) 樣本收集區過濾測試；(4)NC 薄膜 (Nitrocellulose) 進行 MMP-9 偵測和 (5) 銀離子強化等功能，以達成快速的檢測 MMP-9，並得到此裝置 MMP-9 的最低偵測極限 是 1.49 ng mL−1。在未來展望方面，我們希望此裝置能夠結合多重因子偵測並達到定點照護的概念。我們相信這個裝置將會對健康醫療，尤其在口腔照護方面，有長足的貢獻，並解決醫療資源短缺地區在牙周病偵測上的不便。

In 2010, 10.79% of the world’s population, about 750 million people, is affected by chronic periodontitis. Productivity loss from periodontitis alone is approximately $53.99 billion each year, causing a huge economic burden around the world. Therefore,the early prediction and treatment of periodontitis is critical in relieving the economic burden and medical expenses in low-resource settings. In this thesis, we developed a lateral flow immunoassay (LFIA) test strip for periodontitis diagnosis, and used the LFIA test strip to detect MMP-9, which is a standard biomarker for periodontitis. We achieved: (1) conjugation of MMP-9 antibody to AuNPs;(2) capture antibody optimization;(3)sample pad filtration;(4)MMP-9 detection by the 1/2 LFIA test strips and(5) sliver enhancement, and derive the limit of detection (LOD) of 1.49 ngmL−1. In the future, we hope this device can be applied in multiplex detection and achieve the concept of point-of-care (POC).We believe this development will be greatly contributed to health care and welfare,especially in dental care,and resource-limited country.