{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T17:07:37Z","timestamp":1790528857364,"version":"4.1.0"},"reference-count":69,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,1,26]],"date-time":"2021-01-26T00:00:00Z","timestamp":1611619200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R35GM119518"],"award-info":[{"award-number":["R35GM119518"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a worldwide crisis with profound effects on both public health and the economy. In order to combat the COVID-19 pandemic, research groups have shared viral genome sequence data through the Global Initiative on Sharing All Influenza Data (GISAID). Over the past year, \u2248290,000 full SARS-CoV-2 proteome sequences have been deposited in the GISAID. Here, we used these sequences to assess the rate of nonsynonymous mutants over the entire viral proteome. Our analysis shows that SARS-CoV-2 proteins are mutating at substantially different rates, with most of the viral proteins exhibiting little mutational variability. As anticipated, our calculations capture previously reported mutations that arose in the first months of the pandemic, such as D614G (Spike), P323L (NSP12), and R203K\/G204R (Nucleocapsid), but they also identify more recent mutations, such as A222V and L18F (Spike) and A220V (Nucleocapsid), among others. Our comprehensive temporal and geographical analyses show two distinct periods with different proteome mutation rates: December 2019 to July 2020 and August to December 2020. Notably, some mutation rates differ by geography, primarily during the latter half of 2020 in Europe. Furthermore, our structure-based molecular analysis provides an exhaustive assessment of SARS-CoV-2 mutation rates in the context of the current set of 3D structures available for SARS-CoV-2 proteins. This emerging sequence-to-structure insight is beginning to illuminate the site-specific mutational (in)tolerance of SARS-CoV-2 proteins as the virus continues to spread around the globe.<\/jats:p>","DOI":"10.3390\/biology10020091","type":"journal-article","created":{"date-parts":[[2021,1,26]],"date-time":"2021-01-26T08:29:16Z","timestamp":1611649756000},"page":"91","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":87,"title":["One Year of SARS-CoV-2: How Much Has the Virus Changed?"],"prefix":"10.3390","volume":"10","author":[{"given":"Santiago","family":"Vilar","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL 33136, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel G.","family":"Isom","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL 33136, USA"},{"name":"Sylvester Comprehensive Cancer Center, University of Miami, Miami, FL 33136, USA"},{"name":"Institute for Data Science and Computing, University of Miami, Coral Gables, FL 33146, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Doherty, P.C. (2021). What have we learnt so far from COVID-19?. Nat. Rev. Immunol., 1\u20132.","DOI":"10.1038\/s41577-021-00498-y"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1823","DOI":"10.1097\/ACM.0000000000003656","article-title":"Emerging From the COVID-19 Crisis With a Stronger Health Care Workforce","volume":"95","author":"Dow","year":"2020","journal-title":"Acad. Med."},{"key":"ref_3","unstructured":"OECD Policy Responses to Coronavirus (COVID-19) (2020, December 15). The Territorial Impact of COVID-19: Managing the Crisis Across Levels of Government. Available online: http:\/\/www.oecd.org\/coronavirus\/policy-responses\/the-territorial-impact-of-covid-19-managing-the-crisis-across-levels-of-government-d3e314e1\/#section-d1e343."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"105730","DOI":"10.1016\/j.envint.2020.105730","article-title":"Airborne transmission of SARS-CoV-2: The world should face the reality","volume":"139","author":"Morawska","year":"2020","journal-title":"Environ. Int."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"109808","DOI":"10.1016\/j.envres.2020.109808","article-title":"Current emerging SARS-CoV-2 pandemic: Potential direct\/indirect negative impacts of virus persistence and related therapeutic drugs on the aquatic compartments","volume":"188","author":"Race","year":"2020","journal-title":"Environ. Res."},{"key":"ref_6","unstructured":"National Institute of Allergy and Infectious Diseases (NIAID) (2020, September 02). COVID-19 Is an Emerging, Rapidly Evolving Situation, Available online: https:\/\/www.niaid.nih.gov\/diseases-conditions\/coronaviruses."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"165878","DOI":"10.1016\/j.bbadis.2020.165878","article-title":"Insights into SARS-CoV-2 genome, structure, evolution, pathogenesis and therapies: Structural genomics approach","volume":"1866","author":"Naqvi","year":"2020","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.jmii.2020.02.001","article-title":"Emerging threats from zoonotic coronaviruses-from SARS and MERS to 2019-nCoV","volume":"53","author":"Lee","year":"2020","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/s41586-020-2008-3","article-title":"A new coronavirus associated with human respiratory disease in China","volume":"579","author":"Wu","year":"2020","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"14004","DOI":"10.1038\/s41598-020-70812-6","article-title":"Genome-wide analysis of SARS-CoV-2 virus strains circulating worldwide implicates heterogeneity","volume":"10","author":"Islam","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1016\/j.cell.2020.06.043","article-title":"Tracking Changes in SARS-CoV-2 Spike: Evidence that D614G Increases Infectivity of the COVID-19 Virus","volume":"182","author":"Korber","year":"2020","journal-title":"Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"104351","DOI":"10.1016\/j.meegid.2020.104351","article-title":"Emergence of genomic diversity and recurrent mutations in SARS-CoV-2","volume":"83","author":"Acman","year":"2020","journal-title":"Infect. Genet. Evol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e24","DOI":"10.1016\/j.jinf.2020.03.058","article-title":"Evolutionary analysis of SARS-CoV-2: How mutation of Non-Structural Protein 6 (NSP6) could affect viral autophagy","volume":"81","author":"Benvenuto","year":"2020","journal-title":"J. Infect."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1186\/s12967-020-02344-6","article-title":"Emerging SARS-CoV-2 mutation hot spots include a novel RNA-dependent-RNA polymerase variant","volume":"18","author":"Pachetti","year":"2020","journal-title":"J. Transl. Med."},{"key":"ref_15","unstructured":"Comandatore, F., Chiodi, A., Gabrieli, P., Biffignandi, G.B., Perini, M., Ramazzotti, M., Ricagno, S., Rimoldi, S.G., Gismondo, M.R., and Micheli, V. (2020). Identification of variable sites in Sars-CoV-2 and their abundance profiles in time. bioRxiv."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Khan, M.I., Khan, Z.A., Baig, M.H., Ahmad, I., Farouk, A.E., Song, Y.G., and Dong, J.J. (2020). Comparative genome analysis of novel coronavirus (SARS-CoV-2) from different geographical locations and the effect of mutations on major target proteins: An in silico insight. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0238344"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Mercatelli, D., and Giorgi, F.M. (2020). Geographic and Genomic Distribution of SARS-CoV-2 Mutations. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.01800"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Patro, P.P., Sathyaseelan, C., Uttamrao, P.P., and Rathinavelan, T. (2020). Global variation in the SARS-CoV-2 proteome reveals the mutational hotspots in the drug and vaccine candidates. bioRxiv.","DOI":"10.1101\/2020.07.31.230987"},{"key":"ref_19","unstructured":"(2020, November 02). GISAID, the Global Initiative on Sharing All Influenza Data. Available online: https:\/\/www.gisaid.org\/."},{"key":"ref_20","unstructured":"(2020, September 02). RCSB Protein Data Bank. Available online: http:\/\/www.rcsb.org\/."},{"key":"ref_21","unstructured":"Python Software Foundation (2020, July 02). Python Language Reference. Available online: http:\/\/www.python.org."},{"key":"ref_22","unstructured":"MathWorks, Inc. (2020, August 02). MATLAB: The Language of Technical Computing: Computation, Visualization, Programming. Available online: https:\/\/www.mathworks.com\/."},{"key":"ref_23","unstructured":"Our World in Data (2020, September 02). Statistics and Research. Coronavirus (COVID-19) Cases. Available online: https:\/\/github.com\/owid\/covid-19-data\/tree\/master\/public\/data."},{"key":"ref_24","unstructured":"(2010). The PyMOL Molecular Graphics System, Version 2.4.0, Schr\u00f6dinger, LLC."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1126\/science.abe8499","article-title":"SARS-CoV-2 D614G variant exhibits efficient replication ex vivo and transmission in vivo","volume":"370","author":"Hou","year":"2020","journal-title":"Science"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Tegally, H., Wilkinson, E., Giovanetti, M., Iranzadeh, A., Fonseca, V., Giandhari, J., Doolabh, D., Pillay, S., San, E.J., and Msomi, N. (2020). Emergence and rapid spread of a new severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) lineage with multiple spike mutations in South Africa. medRxiv.","DOI":"10.1101\/2020.12.21.20248640"},{"key":"ref_27","unstructured":"ECDC (2020). Rapid Increase of a SARS-CoV-2 Variant with Multiple Spike Protein Mutations Observed in the United Kingdom European Centre for Disease Prevention and Control, ECDC."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"495","DOI":"10.2471\/BLT.20.253591","article-title":"Variant analysis of SARS-CoV-2 genomes","volume":"98","author":"Koyama","year":"2020","journal-title":"Bull. World Health Organ."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1186\/s12929-018-0449-x","article-title":"Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus","volume":"25","author":"Hsin","year":"2018","journal-title":"J. Biomed. Sci."},{"key":"ref_30","unstructured":"Ayub, M.I. (2020). Reporting Two SARS-CoV-2 Strains Based on A Unique Trinucleotide-Bloc Mutation and Their Potential Pathogenic Difference. Preprints."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Hodcroft, E.B., Zuber, M., Nadeau, S., Comas, I., Gonz\u00e1lez Candelas, F., Stadler, T., and Neher, R.A. (2020). Emergence and spread of a SARS-CoV-2 variant through Europe in the summer of 2020. medRxiv.","DOI":"10.1101\/2020.10.25.20219063"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Issa, E., Merhi, G., Panossian, B., Salloum, T., and Tokajian, S. (2020). SARS-CoV-2 and ORF3a: Non-Synonymous Mutations and Polyproline Regions. bioRxiv.","DOI":"10.1101\/2020.03.27.012013"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"27456","DOI":"10.1073\/pnas.2008590117","article-title":"Seasonality and uncertainty in global COVID-19 growth rates","volume":"117","author":"Merow","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_34","first-page":"e13525","article-title":"SARS-CoV-2 viral spike G614 mutation exhibits higher case fatality rate","volume":"74","author":"Cardozo","year":"2020","journal-title":"Int. J. Clin. Pract."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1075","DOI":"10.1038\/s10038-020-0808-9","article-title":"SARS-CoV-2 genomic variations associated with mortality rate of COVID-19","volume":"65","author":"Toyoshima","year":"2020","journal-title":"J. Hum. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","article-title":"Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing","volume":"57","author":"Benjamini","year":"1995","journal-title":"J. R. Stat. Soc. Ser. B"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Maloy, S., and Hughes, K. (2013). Reverse Mutation. Brenner\u2019s Encyclopedia of Genetics, Academic Press. [2nd ed.].","DOI":"10.1016\/B978-0-12-374984-0.01330-9"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/bs.aivir.2019.08.002","article-title":"Chapter Four\u2014Structural insights into coronavirus entry","volume":"Volume 105","author":"Rey","year":"2019","journal-title":"Advances in Virus Research"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.cell.2020.02.058","article-title":"Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein","volume":"181","author":"Walls","year":"2020","journal-title":"Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1038\/s41401-020-0485-4","article-title":"Structural and functional properties of SARS-CoV-2 spike protein: Potential antivirus drug development for COVID-19","volume":"41","author":"Huang","year":"2020","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1007\/s12038-020-00040-7","article-title":"Global efforts on vaccines for COVID-19: Since, sooner or later, we all will catch the coronavirus","volume":"45","author":"Mukherjee","year":"2020","journal-title":"J. Biosci."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Rowe, J.B., Kapolka, N.J., Taghon, G.J., Morgan, W.M., and Isom, D.G. (2020). The evolution and mechanism of GPCR proton sensing. J. Biol. Chem.","DOI":"10.1074\/jbc.RA120.016352"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"5702","DOI":"10.1073\/pnas.1417888112","article-title":"Buried ionizable networks are an ancient hallmark of G protein-coupled receptor activation","volume":"112","author":"Isom","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1021\/acs.biochem.5b00901","article-title":"Regulation of Ras Paralog Thermostability by Networks of Buried Ionizable Groups","volume":"55","author":"Isom","year":"2016","journal-title":"Biochemistry"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/j.molcel.2013.07.012","article-title":"Protons as second messenger regulators of G protein signaling","volume":"51","author":"Isom","year":"2013","journal-title":"Mol. Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1038\/ncb2678","article-title":"Virus entry: What has pH got to do with it?","volume":"15","author":"Helenius","year":"2013","journal-title":"Nat. Cell Biol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1038\/s41422-020-0366-x","article-title":"Mining of epitopes on spike protein of SARS-CoV-2 from COVID-19 patients","volume":"30","author":"Zhang","year":"2020","journal-title":"Cell Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1007\/s11262-005-0070-4","article-title":"Amino acids 1 to 422 of the spike protein of SARS associated coronavirus are required for induction of cyclooxygenase-2","volume":"33","author":"Liu","year":"2006","journal-title":"Virus Genes"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Moreira, R.A., Guzman, H.V., Boopathi, S., Baker, J.L., and Poma, A.B. (2020). Characterization of Structural and Energetic Differences between Conformations of the SARS-CoV-2 Spike Protein. Materials, 13.","DOI":"10.1101\/2020.11.01.363499"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"16409","DOI":"10.1039\/D0NR03969A","article-title":"Quantitative determination of mechanical stability in the novel coronavirus spike protein","volume":"12","author":"Moreira","year":"2020","journal-title":"Nanoscale"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1016\/j.bbrc.2020.04.136","article-title":"Biochemical characterization of SARS-CoV-2 nucleocapsid protein","volume":"527","author":"Zeng","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Ahmed, S.F., Quadeer, A.A., and McKay, M.R. (2020). Preliminary Identification of Potential Vaccine Targets for the COVID-19 Coronavirus (SARS-CoV-2) Based on SARS-CoV Immunological Studies. Viruses, 12.","DOI":"10.1101\/2020.02.03.933226"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"11613","DOI":"10.1073\/pnas.1811345115","article-title":"The mechanism of resistance to favipiravir in influenza","volume":"115","author":"Goldhill","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.antiviral.2011.10.012","article-title":"Identification of a novel resistance mutation for benzimidazole inhibitors of the HCV RNA-dependent RNA polymerase","volume":"93","author":"Delang","year":"2012","journal-title":"Antivir. Res."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"e9492","DOI":"10.7717\/peerj.9492","article-title":"Identification of novel mutations in RNA-dependent RNA polymerases of SARS-CoV-2 and their implications on its protein structure","volume":"8","author":"Chand","year":"2020","journal-title":"PeerJ"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"E3900","DOI":"10.1073\/pnas.1323705111","article-title":"One severe acute respiratory syndrome coronavirus protein complex integrates processive RNA polymerase and exonuclease activities","volume":"111","author":"Subissi","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1016\/j.jmb.2005.09.007","article-title":"Mutational analysis of the SARS virus Nsp15 endoribonuclease: Identification of residues affecting hexamer formation","volume":"353","author":"Guarino","year":"2005","journal-title":"J. Mol. Biol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"3718","DOI":"10.1038\/s41467-020-17496-8","article-title":"Structural basis of RNA cap modification by SARS-CoV-2","volume":"11","author":"Viswanathan","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"10900","DOI":"10.1128\/JVI.00385-06","article-title":"Construction of a severe acute respiratory syndrome coronavirus infectious cDNA clone and a replicon to study coronavirus RNA synthesis","volume":"80","author":"Dediego","year":"2006","journal-title":"J. Virol."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Bz\u00f3wka, M., Mitusi\u0144ska, K., Raczy\u0144ska, A., Samol, A., Tuszy\u0144ski, J.A., and G\u00f3ra, A. (2020). Structural and Evolutionary Analysis Indicate That the SARS-CoV-2 Mpro Is a Challenging Target for Small-Molecule Inhibitor Design. Int. J. Mol. Sci., 21.","DOI":"10.1101\/2020.02.27.968008"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1038\/s41586-020-2223-y","article-title":"Structure of Mpro from SARS-CoV-2 and discovery of its inhibitors","volume":"582","author":"Jin","year":"2020","journal-title":"Nature"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1331","DOI":"10.1126\/science.abb4489","article-title":"Structure-based design of antiviral drug candidates targeting the SARS-CoV-2 main protease","volume":"368","author":"Dai","year":"2020","journal-title":"Science."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.jgg.2020.02.001","article-title":"Potential inhibitors against 2019-nCoV coronavirus M protease from clinically approved medicines","volume":"47","author":"Liu","year":"2020","journal-title":"J. Genet. Genomics"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.str.2004.01.016","article-title":"The nsp9 replicase protein of SARS-coronavirus, structure and functional insights","volume":"12","author":"Sutton","year":"2004","journal-title":"Structure"},{"key":"ref_65","first-page":"814","article-title":"Crystal structures of SARS-CoV-2 ADP-ribose phosphatase: From the apo form to ligand complexes","volume":"7","author":"Michalska","year":"2020","journal-title":"bioRxiv"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.antiviral.2014.12.015","article-title":"The SARS-coronavirus papain-like protease: Structure, function and inhibition by designed antiviral compounds","volume":"115","author":"Mesecar","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Rut, W., Lv, Z., Zmudzinski, M., Patchett, S., Nayak, D., Snipas, S.J., El Oualid, F., Huang, T.T., Bekes, M., and Drag, M. (2020). Activity profiling and structures of inhibitor-bound SARS-CoV-2-PLpro protease provides a framework for anti-COVID-19 drug design. bioRxiv.","DOI":"10.1101\/2020.04.29.068890"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"15189","DOI":"10.1128\/JVI.79.24.15189-15198.2005","article-title":"The Papain-Like Protease of Severe Acute Respiratory Syndrome Coronavirus Has Deubiquitinating Activity","volume":"79","author":"Barretto","year":"2005","journal-title":"J. Virol."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1093\/jalm\/jfaa089","article-title":"Clinical Validation of a SARS-CoV-2 Real-Time Reverse Transcription PCR Assay Targeting the Nucleocapsid Gene","volume":"5","author":"SoRelle","year":"2020","journal-title":"J. Appl. Lab. Med."}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/10\/2\/91\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:15:22Z","timestamp":1760159722000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/10\/2\/91"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,26]]},"references-count":69,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,2]]}},"alternative-id":["biology10020091"],"URL":"https:\/\/doi.org\/10.3390\/biology10020091","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.12.16.423071","asserted-by":"object"}]},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,26]]}}}