Isolation and characterization of an early SARS-CoV-2 isolate from the 2020 epidemic in Medellín, Colombia

Francisco J. Díaz, Wbeimar Aguilar-Jiménez, Lizdany Flórez-Álvarez, Gladys Valencia, Katherine Laiton-Donato, Carlos Franco-Muñoz, Diego Álvarez-Díaz, Marcela Mercado-Reyes, María Teresa Rugeles, .

Keywords: Coronavirus infections, viral isolation severe acute respiratory syndrome, SARS virus, high-throughput nucleotide sequencing, microscopy, electron, fluorescent antibody technique, indirect

Abstract

Introduction: SARS-CoV-2 has been identified as the new coronavirus causing an outbreak of acute respiratory disease in China in December, 2019. This disease, currently named COVID-19, has been declared as a pandemic by the World Health Organization (WHO). The first case of COVID-19 in Colombia was reported on March 6, 2020. Here we characterize an early SARS-CoV-2 isolate from the pandemic recovered in April, 2020.
Objective: To describe the isolation and characterization of an early SARS-CoV-2 isolate from the epidemic in Colombia.
Materials and methods: A nasopharyngeal specimen from a COVID-19 positive patient was inoculated on different cell lines. To confirm the presence of SARS-CoV-2 on cultures we used qRT-PCR, indirect immunofluorescence assay, transmission and scanning electron microscopy, and next-generation sequencing.
Results: We determined the isolation of SARS-CoV-2 in Vero-E6 cells by the appearance of the cytopathic effect three days post-infection and confirmed it by the positive results in the qRT-PCR and the immunofluorescence with convalescent serum. Transmission and scanning electron microscopy images obtained from infected cells showed the presence of structures compatible with SARS-CoV-2. Finally, a complete genome sequence obtained by next-generation sequencing allowed classifying the isolate as B.1.5 lineage.
Conclusion: The evidence presented in this article confirms the first isolation of SARSCoV-2 in Colombia. In addition, it shows that this strain behaves in cell culture in a similar way to that reported in the literature for other isolates and that its genetic composition is consistent with the predominant variant in the world. Finally, points out the importance of viral isolation for the detection of neutralizing antibodies, for the genotypic and phenotypic characterization of the strain and for testing compounds with antiviral potential.

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References

Huang D, Lian X, Song F, Ma H, Lian Z, Liang Y, et al. Clinical features of severe patients infected with 2019 novel coronavirus: A systematic review and meta-analysis. Ann Transl Med. 2020;8:576. https://doi.org/10.21037/atm-20-2124

Wu F, Zhao S, Yu B, Chen YM, Wang W, Song ZG, et al. A new coronavirus associated with human respiratory disease in China. Nature. 2020;579:265-9. https://doi.org/10.1038/s41586-020-2008-3

Zhou P, Yang XL, Wang XG, Hu B, Zhang L, Zhang W, et al. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 2020;579:270-3. https://doi.org/10.1038/s41586-020-2012-7

Zhu N, Zhang D, Wang W, Li X, Yang B, Song J, et al. A novel coronavirus from patients with pneumonia in China, 2019. N Engl J Med. 2020;382:727-33. https://doi.org/10.1056/NEJMoa2001017

Bai Y, Jiang D, Lon JR, Chen X, Hu M, Lin S, et al. Comprehensive evolution and molecular characteristics of a large number of SARS-CoV-2 genomes reveal its epidemic trends. Int J Infect Dis. 2020;100:164-73. https://doi.org/10.1016/j.ijid.2020.08.066

Koyama T, Platt D, Parida L. Variant analysis of SARS-CoV-2 genomes. Bull World Health Organ. 2020;98:495-504. https://doi.org/10.2471/BLT.20.253591

Korber B, Fischer WM, Gnanakaran S, Yoon H, Theiler J, Abfalterer W, et al. Tracking changes in SARS-CoV-2 spike: Evidence that D614G increases infectivity of the COVID-19 virus. Cell. 2020;182:812-27 e19. https://doi.org/10.1016/j.cell.2020.06.043

Toyoshima, Y., Nemoto, K., Matsumoto, S. et al. SARS-CoV-2 genomic variations associated with mortality rate of COVID-19. J Hum Genet. 2020. https://doi.org/10.1038/s10038-020-0808-9

Ministerio de Salud y Protección Social. Colombia confirma su primer caso de COVID-19. Fecha de consulta: 25 de septiembre de 2020. Disponible en: https://www.minsalud.gov.co/Paginas/Colombia-confirma-su-primer-caso-de-COVID-19.aspx

Corman VM, Landt O, Kaiser M, Molenkamp R, Meijer A, Chu DK, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25:2000045. https://doi.org/10.2807/1560-7917.ES.2020.25.3.2000045

World Health Organization. Laboratory biosafety guidance related to coronavirus disease (COVID-19). (COVID-19): Interim guidance, March 19, 2020. Geneva: WHO; 2020. p. 11. Fecha de consulta: 25 de septiembre de 2020. Disponible en: https://apps.who.int/iris/handle/10665/331500?locale-attribute=es&

Quick J. nCoV-2019 sequencing protocol V.1 protocols.io2020. Fecha de consulta: 25 de septiembre de 2020. Disponible en: https://www.protocols.io/view/ncov-2019-sequencingprotocol-bbmuik6w

Rambaut A, Quick J. artic-ncov2019 GitHub2020. Fecha de consulta: 25 de septiembre de 2020. Disponible en: https://github.com/artic-network/artic-ncov2019/blob/master/primer_schemes/nCoV-2019/V1/nCoV-2019_SMARTplex.tsv

Li H, Wong C, Mori Y. bwa - Burrows- Wheeler Alignment Tool 2020. Fecha de consulta: 25 de septiembre de 2020. Disponible en: http://bio-bwa.sourceforge.net/bwa.shtml

O´Toole A, McCrone J. Phylogenetic assignment of named global outbreak LINeages 2020. Fecha de consulta: 18 de junio de 2020. Disponible en: https://github.com/hCoV-2019/pangolin

Laiton-Donato K, Villabona-Arenas CJ, Usme-Ciro JA, Franco-Muñoz C, Álvarez-Díaz DA, Villabona-Arenas LS, et al. Genomic epidemiology of SARS-CoV-2 in Colombia. medRxiv. 2020;medRxiv 2020.06.26.20135715. https://doi.org/10.1101/2020.06.26.20135715

Park WB, Kwon NJ, Choi SJ, Kang CK, Choe PG, Kim JY, et al. Virus isolation from the first patient with SARS-CoV-2 in Korea. J Korean Med Sci. 2020;35:e84. https://doi.org/10.3346/jkms.2020.35.e84

Harcourt J, Tamin A, Lu X, Kamili S, Sakthivel SK, Murray J, et al. Isolation and characterization of SARS-CoV-2 from the first US COVID-19 patient. bioRxiv. 2020;bioRxiv 2020.03.02.972935. https://doi.org/10.1101/2020.03.02.972935

Centers for Disease Control and Prevention. Duration of isolation and precautions for adults with COVID-19 Healthcare workers 2020. Fecha de consulta: 25 de septiembre de 2020. Disponible en: https://www.cdc.gov/coronavirus/2019-ncov/hcp/duration-isolation.html

Zhang L, Jackson CB, Mou H, Ojha A, Rangarajan ES, Izard T, et al. The D614G mutation in the SARS-CoV-2 spike protein reduces S1 shedding and increases infectivity. bioRxiv. 2020;bioRxiv 2020.06.12.14872. https://doi.org/10.1101/2020.06.12.148726

Plante JA, Liu Y, Liu J, Xia H, Johnson BA, Lokugamage KG, et al. Spike mutation D614G alters SARS-CoV-2 fitness and neutralization susceptibility. bioRxiv. 2020;bioRxiv 2020.09.01.278689. https://doi.org/10.1101/2020.09.01.278689

Bullard J, Dust K, Funk D, Strong JE, Alexander D, Garnett L, et al. Predicting infectious severe acute respiratory syndrome coronavirus 2 from diagnostic samples. Clin Infect Dis. 2020. https://doi.org/10.1093/cid/ciaa638

How to Cite
1.
Díaz FJ, Aguilar-Jiménez W, Flórez-Álvarez L, Valencia G, Laiton-Donato K, Franco-Muñoz C, et al. Isolation and characterization of an early SARS-CoV-2 isolate from the 2020 epidemic in Medellín, Colombia. biomedica [Internet]. 2020 Oct. 30 [cited 2024 May 16];40(Supl. 2):148-5. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/5834

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2020-10-30

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