Development of the COVID-19 Real-Time RT-PCR Testing System

Shuryaeva AK1, Malova TV1, Davydova EE2, Savochkina YuA1, Bogoslovskaya EV1, Mintaev RR1,2, Tsyganova GM1, Shivlyagina EE1, Ibragimova ASh1, Nosova AO1, Shipulin GA1, Yudin SM1
About authors

1 Federal State Budgetary Institution "Centre for Strategic Planning and Management of Biomedical Health Risks" of the Federal Medical Biological Agency, Moscow, Russia

2 I. Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia

About paper

Funding: the study received funding from the Strategic Planning Center, Federal State Budgetary Institution under the Federal Medical-Biological Agency of Russia

Received: 2020-07-15 Accepted: 2020-08-13 Published online: 2020-08-19

Late in December 2019, an outbreak of an unknown coronavirus, later identified as SARS-CoV- 2, emerged in the city of Wuhan, China. It causes a dangerous respiratory coronavirus disease in humans - COVID-19.
Objective. To detect cases of the disease and prevent its spread across the Russian Federation it is necessary to create an effective diagnostic test system.
Material and methods.
Based on the analysis of the alignment of the SARS-CoV-2 nucleotide sequences, primers and a probe for RT-PCR were selected, and the analysis conditions were optimized.
Results. The diagnostic system was developed and registered in the shortest possible time in real-time RT-PCR format for detecting SARS-CoV-2 coronavirus RNA in smears from the nasopharynx and oropharynx, sputum and feces. The high specificity of the system was verified on a representative set of viruses and microorganisms, the analytical sensitivity was 1x103 copies / ml in smears from the mucous membrane of the nasopharynx and oropharynx and sputum, 5x104 copies / ml in fecal samples. Diagnostic sensitivity and specificity established during clinical trials on samples from patients with confirmed COVID-19 infection, from patients with a different etiology of a disease and clinically healthy people were to 100% (range 94.2-100% with a confidence level of 95%).

Keywords: coronavirus, molecular diagnostics, COVID-19, SARS-CoV-2, SARS-COV-2, real-time RT-PCR, diagnosis of infectious diseases