利用与适体偶联的产离子酶的电化学生物传感系统用于检测严重急性呼吸综合征冠状病毒2 。

Electrical biosensing system utilizing ion-producing enzymes conjugated with aptamers for the sensing of severe acute respiratory syndrome coronavirus 2.

作者信息

Nukazuka Akira, Asai Satomi, Hayakawa Kei, Nakagawa Kazuhisa, Kanazashi Mana, Kakizoe Hidefumi, Hayashi Kyoko, Kawahara Toshio, Sawada Kazuaki, Kuno Hitoshi, Kano Kazuhiko

机构信息

DENSO Corporation, 1-1 Showa-cho, Kariya, Aichi 448-8661, Japan.

Department of Laboratory Medicine, Tokai University School of Medicine, 143 Shimogasuya, Isehara, Kanagawa 259-1193, Japan.

出版信息

Sens Biosensing Res. 2023 Feb;39:100549. doi: 10.1016/j.sbsr.2023.100549. Epub 2023 Jan 18.

Abstract

Viral outbreaks, which include the ongoing coronavirus disease 2019 (COVID-19) pandemic provoked by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are a major global crisis that enormously threaten human health and social activities worldwide. Consequently, the rapid and repeated treatment and isolation of these viruses to control their spread are crucial to address the COVID-19 pandemic and future epidemics of novel emerging viruses. The application of cost-efficient, rapid, and easy-to-operate detection devices with miniaturized footprints as a substitute for the conventional optic-based polymerase chain reaction (PCR) and immunoassay tests is critical. In this context, semiconductor-based electrical biosensors are attractive sensing platforms for signal readout. Therefore, this study aimed to examine the electrical sensing of patient-derived SARS-CoV-2 samples by harnessing the activity of DNA aptamers directed against spike proteins on viral surfaces. We obtained rapid and sensitive virus detection beyond the Debye length limitation by exploiting aptamers coupled with alkaline phosphatases, which catalytically generate free hydrogen ions which can readily be measured on pH meters or ion-sensitive field-effect transistors. Furthermore, we demonstrated the detection of the viruses of approximately 100 copies/μL in 10 min, surpassing the capability of typical immunochromatographic assays. Therefore, our newly developed technology has great potential for point-of-care testing not only for SARS-CoV-2, but also for other types of pathogens and biomolecules.

摘要

病毒爆发,包括由严重急性呼吸综合征冠状病毒2(SARS-CoV-2)引发的持续的2019冠状病毒病(COVID-19)大流行,是一场重大的全球危机,对全球人类健康和社会活动构成巨大威胁。因此,快速且反复地检测和隔离这些病毒以控制其传播,对于应对COVID-19大流行和新型病毒未来的流行至关重要。应用具有成本效益、快速且易于操作、占地面积小的检测设备来替代传统的基于光学的聚合酶链反应(PCR)和免疫分析测试至关重要。在这种背景下,基于半导体的电化学生物传感器是用于信号读出的有吸引力的传感平台。因此,本研究旨在通过利用针对病毒表面刺突蛋白的DNA适配体的活性,来检测患者来源的SARS-CoV-2样本的电传感。我们通过利用与碱性磷酸酶偶联的适配体,获得了超越德拜长度限制的快速且灵敏的病毒检测,碱性磷酸酶催化产生可在pH计或离子敏感场效应晶体管上轻松测量的游离氢离子。此外,我们证明了在10分钟内可检测到约100拷贝/μL的病毒,超过了典型免疫层析测定的能力。因此,我们新开发的技术不仅对SARS-CoV-2,而且对其他类型的病原体和生物分子的即时检测都具有巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2133/9847365/532a1973b87a/gr1_lrg.jpg

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