FRET-Based haCE2 Receptor Mimic Peptide Conjugated Nanoprobe for Simple Detection of SARS-CoV-2

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has led to a pandemic of acute respiratory disease, namely coronavirus disease (COVID-19). This disease threatens human health and public safety. Early diagnosis, isolation, and prevention are important to suppress the outbreak of COVID 19 given the lack of specific antiviral drugs to treat this disease and the emergence of various variants of the virus that cause breakthrough infections even after vaccine administration. Simple and prompt testing is paramount to preventing further spread of the virus. However, current testing methods, namely RT-PCR, is time-consuming. Binding of the SARS-CoV-2 spike (S) glycoprotein to human angiotensin-converting enzyme 2 (hACE2) receptor plays a pivotal role in host cell entry. In the present study, we developed a hACE2 mimic peptide beacon (COVID19-PEB) for simple detection of SARS-CoV-2 using a fluorescence resonance energy transfer system. COVID19-PEB exhibits minimal fluorescence in its ‘‘closed’’ hairpin structure; however, in the presence of SARS-CoV-2, the specific recognition of the S protein receptor-binding domain by COVID19-PEB causes the beacon to assume an ‘‘open’’ structure that emits strong fluorescence. COVID19-PEB can detect SARS-CoV-2 within 3 h or even 50 min and exhibits strong fluorescence even at low viral concentrations, with a detection limit of 4 × 10 3 plaque-forming unit/test. Furthermore, in SARS-CoV-2-infected patient samples confirmed using polymerase chain reaction, COVID19-PEB accurately detected the virus. COVID19-PEB could be developed as a rapid and accurate diagnostic tool for COVID-19.

Note:
Funding: This research was supported by the Center for BioNano Health Guard funded by the MSIT as a Global Frontier Project (HGUARD_2013M3A6B2078950 and HGUARD_2014M3A6B2060507), the Bio & Medical Technology Development Programs (No. NRF-2018M3A9E2022821 and NRF-2021M3E5E3080379), Basic Science Research Program (NRF-2019R1C1C1006867) of the National Research Foundation (NRF) funded by MSIT, Technology Development Program for Biological Hazards Management in Indoor Air through Korea Environment Industry & Technology Institute (KEITI) funded by Ministry of Environment (ME) of Korea (2021003370003), Industrial Technology Alchemist Program of the Ministry of Trade, Industry, and Energy (MOTIE) of Korea (20012435), the National Research Council of Science & Technology (NST) grant by MSIT (No. CAP-20-897 01- KRIBB), and the KRIBB Research Initiative Program (1711134038 and 1711134081).

Declaration of Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Keywords: COVID-19, SARS-CoV-2, spike protein RBD, hACE2 mimic peptide, FRET probe