Molecular methods for diagnosing novel coronavirus infection: comparison of loop-mediated isothermal amplification and polymerase chain reaction

V. G. Akimkin; Акимкин В. Г.; V. V. Petrov; Петров В. В.; K. V. Krasovitov; Красовитов К. В.; N. I. Borisova; Борисова Н. И.; I. A. Kotov; Котов И. А.; E. N. Rodionova; Родионова Е. Н.; A. S. Cherkashina; Черкашина А. С.; L. Yu. Kondrasheva; Кондрашева Л. Ю.; E. V. Tivanova; Тиванова Е. В.; K. F. Khafizov; Хафизов К. Ф.

doi:10.36233/0507-4088-86

Molecular methods for diagnosing novel coronavirus infection: comparison of loop-mediated isothermal amplification and polymerase chain reaction

作者: Akimkin V.G.¹, Petrov V.V.¹, Krasovitov K.V.¹, Borisova N.I.¹, Kotov I.A.¹, Rodionova E.N.¹, Cherkashina A.S.¹, Kondrasheva L.Y.¹, Tivanova E.V.¹, Khafizov K.F.¹
隶属关系:
1. FSBI «Central Research Institute for Epidemiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
期: 卷 66, 编号 6 (2021)
页面: 417-424
栏目: ORIGINAL RESEARCH
URL: https://ogarev-online.ru/0507-4088/article/view/118199
DOI: https://doi.org/10.36233/0507-4088-86
ID: 118199

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Introduction. Currently, the basis for molecular diagnostics of most infections is the use of reverse transcription polymerase chain reaction (RT-PCR). Technologies based on reverse transcription isothermal loop amplification (RT-LAMP) can be used as an alternative to RT-PCR for diagnostic purposes. In this study, we compared the RTLAMP and RT-PCR methods in order to analyze both the advantages and disadvantages of the two approaches.

Material and methods. For the study, we used reagent kits based on RT-PCR and RT-LAMP. The biological material obtained by taking swabs from the mucous membrane of the oropharynx and nasopharynx in patients with symptoms of a new coronavirus infection was used.

Results. We tested 381 RNA samples of the SARS-CoV-2 virus (Coronaviridae: Coronavirinae: Betacoronavirus; Sarbecovirus) from various patients. The obtained values of the threshold cycle (Ct) for RT-PCR averaged 20.0 ± 3.7 s (1530 ± 300 s), and for RT-LAMP 12.8 ± 3.7 s (550 ± 160 s). Proceeding from the theoretical assumptions, a linear relationship between values obtained in two kits was proposed as a hypothesis; the correlation coefficient was approximately 0.827. At the same time, for samples with a low viral load (VL), the higher Ct values in RT-LAMP did not always correlated with those obtained in RT-PCR.

Discussion. We noted a significant gain in time for analysis using RT-LAMP compared to RT-PCR, which can be important in the context of testing a large number of samples. Being easy to use and boasting short turnaround time, RT-LAMP-based test systems can be used for mass screening in order to identify persons with medium and high VLs who pose the greatest threat of the spread of SARS-CoV-2, while RT-PCR-based diagnostic methods are also suitable for estimation of VL and its dynamics in patients with COVID-19.

关键词

coronavirus infection, coronavirus SARS-CoV-2, polymerase chain reaction (PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP)