A Multiallelic Molecular Beacon-Based Real-Time RT-PCR Assay for the Detection of SARS-CoV-2.
Authors
Hezka Rodosthenous, Johana
Topcu, Cicek
Papa, Christina
Aristokleous, Antonia
Stathi, Georgia
Eleftheriou, Christina
Stylianou, Dora C
Publication Date
2021-10-27Journal Title
Life (Basel)
ISSN
2075-1729
Publisher
MDPI AG
Volume
11
Issue
11
Language
en
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Chrysostomou, A. C., Hezka Rodosthenous, J., Topcu, C., Papa, C., Aristokleous, A., Stathi, G., Christodoulou, C., et al. (2021). A Multiallelic Molecular Beacon-Based Real-Time RT-PCR Assay for the Detection of SARS-CoV-2.. Life (Basel), 11 (11) https://doi.org/10.3390/life11111146
Description
Funder: European Regional Development Fund and the Republic of Cyprus through the Cyprus Research and Innovation Foundation and the Structural Funds 2021-2027; Grant(s): Project: CONCEPT-COVID_0420_0024
Abstract
Emerging infectious viruses have led to global advances in the development of specific and sensitive detection techniques. Viruses have an inherent potential to easily mutate, presenting major hurdles for diagnostics and requiring methods capable of detecting genetically diverse viral strains. One such infectious agent is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged in December 2019 and has resulted in the global coronavirus disease 2019 (COVID-19) pandemic. This study presents a real-time reverse transcription PCR (RT-PCR) detection assay for SARS-CoV-2, taking into account its intrinsic polymorphic nature that arises due to genetic drift and recombination, as well as the possibility of continuous and multiple introductions of genetically nonidentical strains into the human population. This advance was achieved by using mismatch-tolerant molecular beacons designed to specifically detect the SARS-CoV-2 S, E, M, and N genes. These were applied to create a simple and reproducible real-time RT-PCR assay, which was validated using external quality control panels (QCMD: CVOP20, WHO: SARS-CoV-2-EQAP-01) and clinical samples. This assay was designed for high target detection accuracy and specificity and can also be readily adapted for the detection of other emerging and rapidly mutating pathogens.
Keywords
COVID-19, SARS-CoV-2, molecular beacons
Sponsorship
University of Cyprus (“Research Saves Lives” campaign)
Identifiers
External DOI: https://doi.org/10.3390/life11111146
This record's URL: https://www.repository.cam.ac.uk/handle/1810/330037
Rights
Licence:
https://creativecommons.org/licenses/by/4.0/
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