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Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA

The newly identified coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causes coronavirus disease 2019 (COVID-19) and has affected over 25 million people worldwide as of August 31, 2020. To aid in the development of diagnostic kits for rapid and sensitive detection of the vi...

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Autores principales: Varlamov, Dmitriy A., Blagodatskikh, Konstantin A., Smirnova, Evgenia V., Kramarov, Vladimir M., Ignatov, Konstantin B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672014/
https://www.ncbi.nlm.nih.gov/pubmed/33251206
http://dx.doi.org/10.3389/fbioe.2020.604793
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author Varlamov, Dmitriy A.
Blagodatskikh, Konstantin A.
Smirnova, Evgenia V.
Kramarov, Vladimir M.
Ignatov, Konstantin B.
author_facet Varlamov, Dmitriy A.
Blagodatskikh, Konstantin A.
Smirnova, Evgenia V.
Kramarov, Vladimir M.
Ignatov, Konstantin B.
author_sort Varlamov, Dmitriy A.
collection PubMed
description The newly identified coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causes coronavirus disease 2019 (COVID-19) and has affected over 25 million people worldwide as of August 31, 2020. To aid in the development of diagnostic kits for rapid and sensitive detection of the virus, we evaluated a combination of polymerase chain reaction (PCR) and isothermal nucleic acid amplification techniques. Here, we compared conventional PCR and loop-mediated isothermal amplification (LAMP) methods with hybrid techniques such as polymerase chain displacement reaction (PCDR) and a newly developed PCR-LAMP method. We found that the hybrid methods demonstrated higher sensitivity and assay reaction rates than those of the classic LAMP and PCR techniques and can be used to for SARS-CoV-2 detection. The proposed methods based on the modern hybrid amplification techniques markedly improve virus detection and, therefore, can be extremely useful in the development of new diagnostic kits.
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spelling pubmed-76720142020-11-26 Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA Varlamov, Dmitriy A. Blagodatskikh, Konstantin A. Smirnova, Evgenia V. Kramarov, Vladimir M. Ignatov, Konstantin B. Front Bioeng Biotechnol Bioengineering and Biotechnology The newly identified coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causes coronavirus disease 2019 (COVID-19) and has affected over 25 million people worldwide as of August 31, 2020. To aid in the development of diagnostic kits for rapid and sensitive detection of the virus, we evaluated a combination of polymerase chain reaction (PCR) and isothermal nucleic acid amplification techniques. Here, we compared conventional PCR and loop-mediated isothermal amplification (LAMP) methods with hybrid techniques such as polymerase chain displacement reaction (PCDR) and a newly developed PCR-LAMP method. We found that the hybrid methods demonstrated higher sensitivity and assay reaction rates than those of the classic LAMP and PCR techniques and can be used to for SARS-CoV-2 detection. The proposed methods based on the modern hybrid amplification techniques markedly improve virus detection and, therefore, can be extremely useful in the development of new diagnostic kits. Frontiers Media S.A. 2020-11-04 /pmc/articles/PMC7672014/ /pubmed/33251206 http://dx.doi.org/10.3389/fbioe.2020.604793 Text en Copyright © 2020 Varlamov, Blagodatskikh, Smirnova, Kramarov and Ignatov. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Varlamov, Dmitriy A.
Blagodatskikh, Konstantin A.
Smirnova, Evgenia V.
Kramarov, Vladimir M.
Ignatov, Konstantin B.
Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA
title Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA
title_full Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA
title_fullStr Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA
title_full_unstemmed Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA
title_short Combinations of PCR and Isothermal Amplification Techniques Are Suitable for Fast and Sensitive Detection of SARS-CoV-2 Viral RNA
title_sort combinations of pcr and isothermal amplification techniques are suitable for fast and sensitive detection of sars-cov-2 viral rna
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672014/
https://www.ncbi.nlm.nih.gov/pubmed/33251206
http://dx.doi.org/10.3389/fbioe.2020.604793
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