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Peptide substrate screening for the diagnosis of SARS-CoV-2 using fluorescence resonance energy transfer (FRET) assay.
| Content Provider | Europe PMC |
|---|---|
| Author | Alhadrami, Hani A. Hassan, Ahmed M. Chinnappan, Raja Al-Hadrami, Hind Abdulaal, Wesam H. Azhar, Esam I. Zourob, Mohammed |
| Abstract | The novel corona (SARS-CoV-2) virus causes a global pandemic, which motivates researchers to develop reliable and effective methods for screening and detection of SARS-CoV-2. Though there are several methods available for the diagnosis of SARS-CoV-2 such as RT-PCR and ELSIA, nevertheless, these methods are time-consuming and may not apply at the point of care. In this study, we have developed a specific, sensitive, quantitative and fast detection method for SARS-CoV-2 by fluorescence resonance energy transfer (FRET) assay. The total extracellular protease proteolytic activity from the virus has been used as the biomarker. The specific peptide sequences from the library of 115 dipeptides were identified via changes in the fluorescence signal. The fluorogenic dipeptide substrates have the fluorophore and a quencher at the N- and the C- terminals, respectively. When the protease hydrolyzes the peptide bond between the two specific amino acids, it leads to a significant increase in the fluorescence signals. The specific fluorogenic peptide (H-d) produces a high fluorescence signal. A calibration plot was obtained from the changes in the fluorescence intensity against the different concentrations of the viral protease. The lowest limit of detection of this method was 9.7 ± 3 pfu/mL. The cross-reactivity of the SARS-CoV-2-specific peptide was tested against the MERS-CoV which does not affect the fluorescence signal. A significant change in the fluorescence signal with patient samples indicates that this FRET-based assay might be applied for the diagnosis of SARS-CoV-2 patients.Graphical abstract |
| Related Links | https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC7990899&blobtype=pdf |
| ISSN | 00263672 |
| Journal | Mikrochimica Acta [Mikrochim Acta] |
| Volume Number | 188 |
| DOI | 10.1007/s00604-021-04766-5 |
| PubMed Central reference number | PMC7990899 |
| Issue Number | 4 |
| PubMed reference number | 33763734 |
| e-ISSN | 14365073 |
| Language | English |
| Publisher | Springer Vienna |
| Publisher Date | 2021-03-25 |
| Publisher Place | Vienna |
| Access Restriction | Open |
| Rights License | This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. © The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature 2021 |
| Subject Keyword | SARS-CoV-2 detection COVID-19 FRET substrate Detection of Covid-19 Fluorogenic peptide sequences |
| Content Type | Text |
| Resource Type | Article |
| Subject | Analytical Chemistry |
