設施農業：智慧感測【生物產業機電工程學系/吳靖宙教授】
| 論文篇名 | 英文：A Label-Free Electrochemical Impedimetric Immunosensor with Biotinylated-Antibody for SARS-CoV-2 Nucleoprotein Detection in Saliva 中文：使用免標定式電化學阻抗免疫感測器對SARS-CoV-2 核殼蛋白的唾液檢測 |
| 期刊名稱 | BIOSENSORS |
| 發表年份,卷數,起迄頁數 | 2022, 12, 265. |
| 作者 | Wu, Ching-Chou(吳靖宙)*; Chiang, Yu-Huan; Chiang, Hsin-Yu |
| DOI | 10.3390/bios12050265 |
| 中文摘要 | 即時性檢測SARS-CoV-2冠狀病毒的抗原對驗證是否感染對控制COVID-19疫情有急迫性的需求。本研究以免標定式電化學阻抗分析(electrochemical impedance spectroscopy, EIS)技術結合鍍金奈米結構之網板印碳電極(AuNS/SPCE)開發能在唾液中檢測SARS-CoV-2核殼蛋白(nucleocapsid protein, N-protein)的免疫感測器 使用短鏈的3-mercaptopropionic acid為自組裝薄膜，再將streptavidin 與bovine serum albumin 共價鍵結其上，隨後可將biotinylated anti-N-protein antibody吸附在此感測膜上，以製作免疫感測器。該感測器具有低的檢測極限(6 pg/mL)，以及良好線性範圍在0.01 ng/mL-100 ng/mL。與MERS-CoV N-protein, influenza A N-protein, influenza B N-protein和 SARS-CoV-2 spike protein之間具有低的交叉反應活性，顯示具有高的專一性。該感測器在健康志願者口水中亦能檢測到6 pg/mL N-protein檢測極限。本研究顯示該拋棄式EIS免疫感測器具有潛力能對唾液中COVID-19進行快速與靈敏的診斷。 |
| 英文摘要 | The timely detecting of SARS-CoV-2 coronavirus antigens for infection validation is an urgent request for COVID-19 pandemic control. This study constructed label-free electrochemical impedance spectroscopy (EIS)-based immunosensors based on gold nanostructured screen-printed carbon electrodes (AuNS/SPCEs) to detect the SARS-CoV-2 nucleocapsid protein (N-protein) in saliva. Using short-chain 3-mercaptopropionic acid (MPA) as a linker to covalently bond streptavidin (SA) and bovine serum albumin (BSA) for controlling the oriented immobilization of the biotinylated anti-N-protein antibody (BioAb) can offer a greater sensitivity, a lower limit of detection (LOD), and better reproducibility of immunosensors (defined as BioAb/SA-BSA/MPA/AuNS/SPCEs) than the antibody randomly immobilized immunosensors and the long-chain 11-mercaptoundecanoic acid (MUA)-modified immunosensors (BioAb/SA-BSA/MUA/AuNS/SPCEs). The BioAb/SA-BSA/MPA/AuNS/SPCE-based immunosensors presented good linearity from 0.01 ng/mL to 100 ng/mL and a low LOD of 6 pg/mL in a phosphate buffer solution (PBS) and PBS-diluted saliva. Moreover, the immunosensor exhibited little cross-activity with other viral antigens such as MERS-CoV N-protein, influenza A N-protein, influenza B N-protein, and SARS-CoV-2 spike protein, indicating the high specificity of the immunosensors. The disposable label-free EIS-based immunosensors have promising potential in facilitating the rapid and sensitive tests of saliva-based COVID-19 diagnostics. |
| 發表成果與本中心研究主題相關性 | 金奈米結構沉積的網版印刷碳電極可製作出高敏感性的免疫感測器，利於應用在環境或農業經濟動物或作物的病毒檢測或藥物殘留分析等領域。 |
