Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences , Wuhan University , Wuhan 430072 , China.
Anal Chem. 2019 Mar 5;91(5):3367-3373. doi: 10.1021/acs.analchem.8b04623. Epub 2019 Feb 20.
Aptamers for Ebola virus (EBOV) offer a powerful means for prevention and diagnostics. Unfortunately, few aptamers for EBOV have been discovered yet. Herein, assisted by magnetism-controlled selection chips to strictly manipulate selection conditions, a highly efficient aptamer selection platform for EBOV is proposed. With highly stringent selection conditions of rigorous washing, manipulation of minuscule amounts of magnetic beads, and real-time evaluation of the selection effectiveness, the selection performance of the platform was improved significantly. In only three rounds of selection, the high-performance aptamers for EBOV GP and NP proteins were obtained simultaneously, with dissociation constants ( K) in the nanomolar range. The aptamer was further applied to the detection of EBOV successfully, with a detection limit of 4.2 ng/mL. The whole detection process that consisted of sample mixing, separation, and signal acquisition was highly integrated and conducted in a magnetism-controlled detection chip, showing high biosafety and great potential for point-of-care detection. The method may open up new avenues for prevention and control of EBOV.
适体(aptamers)作为埃博拉病毒(Ebola virus,EBOV)的有力预防和诊断工具,目前仅有少数适体被发现。本研究提出了一种基于磁控筛选芯片的埃博拉病毒适体高效筛选平台。通过严格的筛选条件(包括严格的洗涤、微量磁珠操作和实时评估筛选效果),显著提高了平台的筛选性能。仅经过三轮筛选,就同时获得了针对埃博拉病毒 GP 和 NP 蛋白的高性能适体,解离常数(K)达到纳摩尔级。该适体还成功应用于埃博拉病毒的检测,检测限为 4.2ng/mL。整个检测过程包括样品混合、分离和信号采集,高度集成并在磁控检测芯片中进行,具有高度的生物安全性和用于即时检测的巨大潜力。该方法可能为埃博拉病毒的预防和控制开辟新途径。