Open AccessMolecular methods for diagnosing novel coronavirus infection: comparison of loop-mediated isothermal amplification and polymerase chain reaction
Author(s) -
В. Г. Акимкин,
Vadim V. Petrov,
Kirill V. Krasovitov,
Nadezhda I. Borisova,
Ivan A. Kotov,
E. N. Rodionova,
A. S. Cherkashina,
L. Yu. Kondrasheva,
Е. В. Тиванова,
K. Khafizov
Publication year - 2022
Publication title -
voprosy virusologii
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.151
H-Index - 15
eISSN - 2411-2097
pISSN - 0507-4088
DOI - 10.36233/0507-4088-86
Subject(s) - reverse transcription loop mediated isothermal amplification , loop mediated isothermal amplification , reverse transcriptase , reverse transcription polymerase chain reaction , real time polymerase chain reaction , polymerase chain reaction , virology , coronavirus , biology , microbiology and biotechnology , medicine , covid-19 , dna , gene , pathology , genetics , messenger rna , disease , infectious disease (medical specialty)
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.