基于双核酸适配体的磁珠-SERS标签三明治结构用于SARS-CoV-2病毒表面S蛋白的快速检测

Double Aptamer-Based Magnetic Bead-SERS-Tagged Sandwich Structure for Rapid Detection of SARS-CoV-2 Virus S Protein

面对新型冠状病毒2019(COVID-19)传播速度极快的情况,发展快速、准确和低成本的诊断方法以检测SARS-CoV-2病毒的特定抗原非常必要。基于逆转录聚合酶链反应(RT-PCR)的COVID-19检测需要特定的实验室,耗时较长。本文针对SARS-CoV-2刺突蛋白,提出了一种基于核酸适配体特异性识别的一步捕获底物和检测探针的表面增强拉曼光谱(SERS)检测平台。利用模拟病毒(PSV)进行实验,不经预处理,在5 min内用便携式拉曼光谱仪检测SARS-CoV-2病毒及其变异株。实验结果表明,针对模拟病毒的检出限为200 TU/mL。此外,该方法可以检测另外5种SARS-CoV-2的变异病毒株,咽拭子体系中的极限检测限可以达到5000 TU/mL。实际大批量咽拭子的实验结果可以达到特异性为100%的效果。因此,该平台在SARS-CoV-2的医护点快速诊断中具有很大的应用潜力。

In the face of the extremely rapid spread of coronavirus disease 2019,it is necessary to develop a rapid,accurate and low-cost diagnostic method to detect specific antigens of SARS-CoV-2 virus.Reverse transcriptase polymerase chain reaction(RT-PCR)-based COVID-19 testing requires specific laboratories and is time-consuming.In this study,we proposed a SERS detection platform for SARS-CoV-2 spike protein based on aptamer specific recognition.This procedure allowed simultaneous capture of the substrate and detection probe.SARS-CoV-2 virus and its variants were detected by portable Raman spectrometer within 5 minutes without pretreatment using simulated virus.The experimental results showed that the detection limit of the simulated virus was 200 TU/mL.In addition,the assay could be used for at least five additional variants of SARS-CoV-2 virus,with a limit line of 5 000 TU/mL in the throat swab system.The results of the actual throat swab experiment could be achieved 100%.Therefore,this platform has great application potential in point-of-care rapid diagnosis of SARS-CoV-2.