Development of a test system and a method for detecting ribonucleic acid of severe acute respiratory syndrome coronavirus 2 using real-time polymerase chain reaction
- Authors: Miteva O.A.1, Smirnova A.V.1, Myasnikova I.A.1, Bykova K.A.1, Kanevsky B.A.1, Myasnikov V.A.1
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Affiliations:
- State Research and Testing Institute of Military Medicine
- Issue: Vol 24, No 4 (2022)
- Pages: 675-682
- Section: Research paper
- Submitted: 05.05.2022
- Accepted: 27.10.2022
- Published: 04.01.2023
- URL: https://journals.eco-vector.com/1682-7392/article/view/107133
- DOI: https://doi.org/10.17816/brmma107133
- ID: 107133
Cite item
Abstract
Given the rapid spread of coronavirus disease 2019 (COVID-19) globally, test systems are needed for its diagnosis, timely treatment, and introduction of quarantine measures. In the shortest possible time, a diagnostic system based on real-time reverse-transcription polymerase chain reaction to detect the ribonucleic acid of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal and oropharyngeal smears was developed and registered. The method determines the nucleocapsid and small-membrane protein genes and the human PGK1 gene, acting as internal control reactions. The nucleotide sequences of SARS-CoV-2 were analyzed, and primers were selected. The conditions for carrying out real-time reverse-transcription polymerase chain reaction and the composition of a set of reagents were set. The diagnostic sensitivity and specificity of the kit were tested on biological samples, with the addition of inactivated SARS-CoV-2. The high analytical characteristics of the developed set of reagents were demonstrated, with a sensitivity of at least 103 GE/mL and a specificity of 100%, and no false-positive or false-negative results were recorded. The high specificity of the test system was shown on a representative sample of genetic materials of respiratory viral pathogens. Clinical and laboratory tests of the diagnostic “SARS-CoV-2 test” were conducted in the N.F. Gamalei National Research Center for Epidemiology and Microbiology. A set of reagents for the detection of ribonucleic acid of SARS-CoV-2 through on real-time reverse-transcription polymerase chain reaction for in vitro diagnostics “SARS-CoV-2 test” was registered in the Russian Federation as a medical device (Registration certificate no. RZN 2020/10632, dated 06/03/2020).
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About the authors
Olga A. Miteva
State Research and Testing Institute of Military Medicine
Author for correspondence.
Email: letto2004@inbox.ru
ORCID iD: 0000-0002-3874-6954
SPIN-code: 2070-7250
Scopus Author ID: 55195685300
applicant for an academic degree
Russian Federation, Saint PetersburgAnna V. Smirnova
State Research and Testing Institute of Military Medicine
Email: gniii_7@mil.ru
SPIN-code: 4897-0219
applicant for an academic degree
Russian Federation, Saint PetersburgIrina A. Myasnikova
State Research and Testing Institute of Military Medicine
Email: gniii_7@mil.ru
SPIN-code: 8883-1534
candidate of medical sciences
Russian Federation, Saint PetersburgKsenia A. Bykova
State Research and Testing Institute of Military Medicine
Email: gniii_7@mil.ru
SPIN-code: 3310-3572
junior research assistant
Russian Federation, Saint PetersburgBoris A. Kanevsky
State Research and Testing Institute of Military Medicine
Email: gniii_7@mil.ru
SPIN-code: 2549-9294
deputy chief
Russian Federation, Saint PetersburgVadim A. Myasnikov
State Research and Testing Institute of Military Medicine
Email: gniii_7@mil.ru
SPIN-code: 5084-2723
candidate of medical sciences
Russian Federation, Saint PetersburgReferences
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