Biosensors Laboratory, Department of Biomedical Engineering, Faculty of Engineering, Mahidol University, Nakhon Pathom, Thailand.
Center of Excellence in Clinical Virology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Nat Commun. 2021 Feb 5;12(1):802. doi: 10.1038/s41467-021-21121-7.
Coronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Diagnosis of COVID-19 depends on quantitative reverse transcription PCR (qRT-PCR), which is time-consuming and requires expensive instrumentation. Here, we report an ultrasensitive electrochemical biosensor based on isothermal rolling circle amplification (RCA) for rapid detection of SARS-CoV-2. The assay involves the hybridization of the RCA amplicons with probes that were functionalized with redox active labels that are detectable by an electrochemical biosensor. The one-step sandwich hybridization assay could detect as low as 1 copy/μL of N and S genes, in less than 2 h. Sensor evaluation with 106 clinical samples, including 41 SARS-CoV-2 positive and 9 samples positive for other respiratory viruses, gave a 100% concordance result with qRT-PCR, with complete correlation between the biosensor current signals and quantitation cycle (Cq) values. In summary, this biosensor could be used as an on-site, real-time diagnostic test for COVID-19.
新型冠状病毒病（COVID-19）是由严重急性呼吸系统综合征冠状病毒 2（SARS-CoV-2）引起的高度传染性疾病。COVID-19 的诊断依赖于耗时且需要昂贵仪器的定量逆转录聚合酶链反应（qRT-PCR）。在此，我们报告了一种基于等温环介导的滚环扩增（RCA）的超灵敏电化学生物传感器，用于快速检测 SARS-CoV-2。该测定法涉及 RCA 扩增子与探针的杂交，探针上功能化有氧化还原活性标记物，可通过电化学生物传感器检测。一步式夹心杂交测定法可以在不到 2 小时的时间内检测到低至 1 拷贝/μL 的 N 和 S 基因。对包括 41 个 SARS-CoV-2 阳性和 9 个其他呼吸道病毒阳性的 106 个临床样本进行传感器评估，与 qRT-PCR 具有 100%的一致性结果，电化学生物传感器的电流信号与定量循环（Cq）值完全相关。总之，该生物传感器可用作 COVID-19 的现场实时诊断测试。
