A simple method for detection of mutations in amino acid 452 of the Spike protein of SARS-CoV-2 using restriction enzyme analysis.
Un método simple para la detección de mutaciones en el aminoácido 452 de la proteína de la Espiga del SARS-CoV-2, usando análisis de enzimas de restricción.
Palabras clave:
COVID-19, SARS-CoV-2, Delta Variant of Concern, RFLP, detección rápida, mutación L452R
Resumen
Las variantes de preocupación o interés del SARS-CoV-2 (VOC o VOI, por sus siglas en inglés), el coronavirus responsable de la COVID-19, han surgido en varios países. Las mutaciones en el aminoácido 452 de la proteína de la Espiga son particularmente importantes y están asociadas con algunas de estas variantes: L452R, presente en la VOC Delta, y L452Q, presente en la VOI Lambda. Estas mutaciones se han asociado con un aumento de la infectividad y la evasión de respuesta inmunitaria protectora. Una búsqueda en GISAID para detectar el número de secuencias que albergan la mutación L452R y la frecuencia de la VOC Delta entre ellas, mostró que desde agosto de 2021, la mayoría de estas secuencias pertenecen a la VOC Delta. Se propone el análisis de enzimas de restricción como un método rápido para detectar L452R. Se digirió un pequeño amplicón de un fragmento del gen de la espiga con MspI. Se observó una concordancia del 100% entre la identificación de la mutación a través de la digestión y los resultados de la secuenciación. La mutación L452Q también se puede detectar mediante análisis de restricción, lo que permite la identificación de posibles VOI Lambda. La metodología propuesta, que permite el cribado de un gran número de muestras, podría contribuir a proporcionar más información sobre la prevalencia y a detectar rápidamente los casos de la VOC Delta.
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Citas
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Phan T. Genetic diversity and evolution of SARS-CoV-2. Infect Genet Evol 2020; 81:104260. Available from: https://doi. org/10.1016/j.meegid.2020.104260.
Rotondo JC, Martini F, Maritati M, Mazziotta C, Di Mauro G, Lanzillotti C, Barp N, Gallerani A, Tognon M, Contini C. SARS-CoV-2 infection: new molecular, phylogenetic, and pathogenetic insights. Efficacy of current vaccines and the potential risk of variants. Viruses 2021; 13:1687. https://doi.org/10.3390/ v13091687.
World Health Organization. https://www. who.int/publications/m/item/weekly-epi- demiological-update-on-covid-19---22-ju- ne-2021.
Noori M, Nejadghaderi SA, Arshi S, Carson-Chahhoud K, Ansarin K, Kolahi AA, Safir S. Potency of BNT162b2 and mRNA-1273 vaccine-induced neutralizing anti-bodies against severe acute respiratory syndrome-CoV-2 variants of concern: A systematic review of in vitro studies. Rev Med Virol 2021;e2277. doi: 10.1002/rmv.2277.
Salleh MZ, Derrick JP, Deris ZZ. Structural evaluation of the spike glycoprotein variants on SARS-CoV-2 transmission and immune rvasion. Int J Mol Sci 2021; 22: 7425. doi: 10.3390/ijms22147425.
European Centre for Disease Prevention and Control. SARS-CoV-2 variants of concern as of 18 June 2021. https://www.ecdc. europa.eu/en/cases-2019-ncov-eueea
Tchesnokovaa V, Kulasekarac H, Larsona L, Bowersd V, Rechkinab E, Kisielaa D, Slednevab Y, Choudhury D, Maslovab I, Dengc K, Kutumbakac K, Gengc H, Fowlerc C, Greenee D, RalstoneJ, Samadpourc M, Sokurenkoa E. Acquisition of the L452R mutation in the ACE2-binding interface of Spike protein triggers recent massive expansion of SARS-CoV-2 variants. J Clin Microbiol 2021:JCM0092121. doi: 10.1128/JCM.00921-21..
Del Rio C, Malani PN, Omer SB. Confronting the Delta Variant of SARS-CoV-2, Summer 2021. JAMA 2021: E1-E2.
Herlihy R, Bamberg W, Burakoff A, Alden N, Severson R, Bush E, Kawasaki B, Berger B, Austin E, Shea M, Gabrieloff E, Matzinger S, Burdorf A, Nichols J, Goode K, Cilwick A, Stacy C, Staples E, Stringer G. Rapid increase in circulation of the SARS-CoV-2B.1.617.2 (Delta) variant—Mesa County, Colorado, April–June 2021. MMWR Morb Mortal Wkly Rep 2021;70:1084–7. https://doi. org/10.15585 /mmwr.mm7032e2
Mishra S, Mindermann S, Sharma M, Whittaker C, Mellan TA, Wilton T, Klapsa D, Mate R, Fritzsche M, Zambon M, Ahuja M, Howes A, Miscouridou X, Nason GP, Ratmann O, Semenova E, Leech G, Sandkühler JF, Rogers-Smith C, Vollmer M Unwin HJT, Gal Y, Chand M, Gandy A, Martin J, Volz E, Ferguson NM, Bhatt S, Brauner JS, Flaxman S, COVID-19 Genomics UK (COG-UK) Consortium. Changing composition of SARS-CoV-2 lineages and rise of Delta variant in England. EClinical- Medicine 2021;39:101064. doi: 10.1016/j. eclinm.2021.101064.
Teyssou E, Delagrèverie H, Visseaux B, Lambert-Niclot S, Brichler S, Ferre V, Marot S, Jary A, Todesco E, Schnuriger A, Ghidaoui E, Abdi B, Akhavan S, Houhou-Fidouh N, Charpentier C, Morand- Joubert L, Boutolleau D, Descamps D, Calvez V, Marcelin AG, Souliea C. The Delta SARS-CoV-2 variant has a higher viral load than the Beta and the historical variants in nasopharyngeal samples from newly diagnosed COVID-19 patients. J Infect 2021; 83:e1-e3. doi: 10.1016/ j.jinf.2021.08.027.
Liu Y, Rocklöv J. The reproductive number of the Delta variant of SARS-CoV-2 is far higher compared to the ancestral SARS- CoV-2 virus. J Travel Med 2021;taab124. doi: 10.1093/jtm/taab124.
Ong SWX, Chiew CJ, Ang LW, Mak T-M, Cui L, Toh MPHS, Lim YD, Lee PH, Lee TH, Chia PY, Maurer-Stroh S, Lin RTP, Leo Y-S, Lee VJ, Lye DC, Young BE. Clinical and virological features of SARS-CoV-2 variants of concern: a retrospective cohort study comparing B.1.1.7 (Alpha), B.1.315 (Beta), and B.1.617.2 (Delta). Clin Infect Dis 2021;ciab721. doi: 10.1093/cid/ ciab721.
Padilla-Rojas C, Jimenez-Vasquez V, Hurtado V, Mestanza O, Molina IS, Barcena L, Morales Ruiz S, Acedo S, Lizarraga W, Bailon H, Cáceres O, Galarza M, Rojas- Serrano N, Vargas-Herrera N, Lope-Pari P, Huayra J, Solari L. Genomic analysis reveals a rapid spread and predominance of lambda (C.37) SARS-COV-2 lineage in Peru despite circulation of variants of concern. J Med Virol 2021:10.1002/jmv.27261.
Jaspe RC, Sulbaran Y, Loureiro CL, D´Angelo P, Rodríguez L, Garzaro DJ, Rangel HR, Pujol FH. Importance of mutations in amino acid 484 of the Spike protein of SARS- CoV-2: rapid detection by restriction enzyme analysis. Invest Clin 2021; 62(Suppl. 2): 3-17.
Wang Y, Chen R, Hu F, Lan Y, Yang Z, Zhan C, Shi J, Deng X, Jiang M, Zhong S, Liao B, Deng K, Tang J, Guo L, Jiang M, Fan Q, Li M, Liu J, Shi Y, Deng X, Xiao X, Kang M, Li Y, Guan W, Li Y, Li S, Li F, Zhong N, Tang X. Transmission, viral kinetics and clinical characteristics of the emergent SARS- CoV-2 Delta VOC in Guangzhou, China. EClinicalMedicine 2021;40:101129. doi: 10.1016/ j.eclinm.2021.101129.
Takeda M. Proteolytic activation of SARS- CoV-2 spike protein. Microbiol Immunol. 2021. doi: 10.1111/1348-0421.12945.
Publicado
2021-11-29
Cómo citar
Jaspe, R. C., Sulbaran, Y., Hidalgo, M., Loureiro, C. L., Moros, Z. C., Garzaro, D. J., Rangel, H. R., & Pujol, F. H. (2021). A simple method for detection of mutations in amino acid 452 of the Spike protein of SARS-CoV-2 using restriction enzyme analysis.: Un método simple para la detección de mutaciones en el aminoácido 452 de la proteína de la Espiga del SARS-CoV-2, usando análisis de enzimas de restricción. Investigación Clínica, 62(4), 371-377. https://doi.org/10.22209/IC.v62n4a07
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