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Mass-Spectrometric Detection of SARS-CoV-2 Virus in Scrapings of the Epithelium of the Nasopharynx of Infected Patients via Nucleocapsid N Protein

Е. Н. Николаев, Maria I. Indeykina, Alexander Brzhozovskiy, Anna E. Bugrova, А. С. Кононихин, Natalia Starodubtseva, Evgeny V. Petrotchenko, Kovalev Gi, Christoph H. Borchers, Г. Т. Сухих

2020Journal of Proteome Research108 citationsDOIOpen Access PDF

Abstract

2020, 119 (6) 1123), and is relatively time-consuming. Fast and reliable complementary methods for detecting the viral infection would be of help in the current pandemic conditions. Mass spectrometry is one of such possibilities. We have developed a mass-spectrometry-based method for the detection of the SARS CoV-2 virus in nasopharynx epithelial swabs based on the detection of the viral nucleocapsid N protein. Our approach shows confident identification of the N protein in patient samples, even those with the lowest viral loads, and a much simpler preparation procedure. Our main protocol consists of virus inactivation by heating and the addition of isopropanol and tryptic digestion of the proteins sedimented from the swabs followed by MS analysis. A set of unique peptides, produced as a result of proteolysis of the nucleocapsid phosphoprotein of SARS-CoV-2, is detected. The obtained results can further be used to create fast parallel mass-spectrometric approaches for the detection of the virus in the nasopharyngeal mucosa, saliva, sputum and other physiological fluids.

Topics & Concepts

Sars virusVirologySevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2)EpitheliumVirusMicrobiologyCoronavirus disease 2019 (COVID-19)Biology2019-20 coronavirus outbreakMedicineOutbreakPathologyDiseaseInfectious disease (medical specialty)GeneticsSARS-CoV-2 and COVID-19 ResearchSARS-CoV-2 detection and testingRespiratory viral infections research