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

Dmitriy A Varlamov; Konstantin A Blagodatskikh; Evgenia V Smirnova; Vladimir M Kramarov; Konstantin B Ignatov

doi:10.3389/fbioe.2020.604793

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

Front Bioeng Biotechnol. 2020 Nov 4:8:604793. doi: 10.3389/fbioe.2020.604793. eCollection 2020.

Authors

Dmitriy A Varlamov¹, Konstantin A Blagodatskikh², Evgenia V Smirnova³, Vladimir M Kramarov⁴, Konstantin B Ignatov^{3

4}

Affiliations

¹ Syntol JSC, Moscow, Russia.
² Laboratory of Molecular Oncology, Pirogov Russian National Research Medical University, Moscow, Russia.
³ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
⁴ Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.

Abstract

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.

Keywords: COVID-19; PCR-LAMP; SARS-CoV-2; hybrid virus detection technique; loop-mediated isothermal amplification; polymerase chain displacement reaction.