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

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 SARSCoV-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). The article can be used under the CC BY-NC-ND 4.0 license © Authors, 2022.

Respiratory RNA Viruses: How to Be Prepared for an Encounter with New Pandemic Virus Strains

The characteristics of the biology of influenza viruses and coronavirus that determine the implementation of the infectious process are presented. With provision for pathogenesis of infection possible effects of serine proteinase inhibitors, heparin, and inhibitors of heparan sulfate receptors in the prevention of cell contamination by viruses are examined. It has been determined that chelators of metals of variable valency and antioxidants should be used for the reduction of replicative activity of viruses and anti-inflammatory therapy. The possibility of a pH-dependent impairment of glycosylation of cellular and viral proteins was traced for chloroquine and its derivatives. The use of low-toxicity drugs as part of adjunct therapy increases the effectiveness of synthetic antiviral drugs and interferons and ensures the safety of baseline therapy.

Respiratory RNA Viruses: How to Prepare for Meeting with New Pandemic Strains

The biology features of influenza viruses and coronaviruses that determine the implementation of the infectious process are described. Taking into account the pathogenesis of infection, the possible effects of serine proteinase blockers, heparin and heparan sulfate receptor blockers in the prevention of cell contamination by viruses are considered. The necessity of using chelators of metals of variable valency and antioxidants to reduce the replicative activity of viruses and anti-inflammatory therapy is determined. The possibility of a pH-dependent violation of the glycosylation of cellular and viral proteins is traced for chloroquine and its derivatives. The use of low-toxic registered drugs as part of adjuvant therapy increases the effectiveness of antiviral synthetic drugs and interferons, ensures the safety of the use of basic therapy. Изложены особенности биологии вирусов гриппа и коронавирусов, определяющие реализацию инфекционного процесса. С учетом патогенеза инфекции рассмотрены возможные эффекты блокаторов сериновых протеиназ, гепарина и блокаторов гепарансульфатных рецепторов в профилактике контаминации клеток вирусами. Определена необходимость применения хелаторов металлов переменной валентности и антиоксидантов для снижения репликативной активности вирусов и противовоспалительной терапии. Возможность pH-зависимого нарушения гликозилирования клеточных и вирусных белков прослежена для хлорохина и его производных. Применение низкотоксичных лекарственных средств в составе вспомогательной терапии повышает эффективность противовирусных синтетических препаратов и интерферонов, обеспечивает безопасность применения средств базисной терапии.