Department Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers, Institution College of Life Sciences, Linyi University, Linyi, 276005, P. R. China.
Chem Asian J. 2022 Nov 2;17(21):e202200747. doi: 10.1002/asia.202200747. Epub 2022 Sep 20.
The main protease (M ), which is highly conserved and plays a critical role in the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a natural biomarker for SARS-CoV-2. Accurate assessment of the M activity is crucial for the detection of SARS-CoV-2. Herein, we report a nanopore-based sensing strategy that uses an enzyme-catalyzed cleavage reaction of a peptide substrate to measure the M activity. The peptide was specifically cleaved by the M , thereby releasing the output products that, when translocated through aerolysin, quantitatively produced the signature current events. The proposed method exhibited high sensitivity, allowing the detection of M concentrations as low as 1 nM without the use of any signal amplification techniques. This simple, convenient, and label-free nanopore assay may expand the diagnostic tools for viruses.
主蛋白酶（M）高度保守，在严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）的复制中起着关键作用，是 SARS-CoV-2 的天然生物标志物。准确评估 M 的活性对于检测 SARS-CoV-2 至关重要。在此，我们报告了一种基于纳米孔的传感策略，该策略使用酶促切割肽底物的反应来测量 M 的活性。该肽被 M 特异性切割，从而释放出产物，当这些产物穿过 aerolysin 时，会定量产生特征电流事件。所提出的方法具有很高的灵敏度，允许检测低至 1 nM 的 M 浓度，而无需使用任何信号放大技术。这种简单、方便、无标记的纳米孔检测方法可能会扩展病毒的诊断工具。
