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Isolation and characterization of an early SARS-CoV-2 isolate from the 2020 epidemic in Medellín, Colombia
Biomedica. 2020 10 30; 40(Supl. 2):148-158.B

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.

Authors+Show Affiliations

Grupo de Inmunovirología, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia. franciscodiaz314@gmail.com.Grupo de Inmunovirología, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia. wbeimar.aguilar@udea.edu.co.Grupo de Inmunovirología, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia. lizdany.florez@udea.edu.co.Ayudas Diagnósticas de Laboratorio Clínico, ADILAB, Medellín, Colombia. gerencia@adilab.com.co.Unidad de Secuenciación Genómica, Instituto Nacional de Salud, Bogotá, D.C., Colombia. klaiton@ins.gov.co.Unidad de Secuenciación Genómica, Instituto Nacional de Salud, Bogotá, D.C., Colombia. cefrancom@unal.edu.co.Unidad de Secuenciación Genómica, Instituto Nacional de Salud, Bogotá, D.C., Colombia. daalvarezd@unal.edu.co.Unidad de Secuenciación Genómica, Instituto Nacional de Salud, Bogotá, D.C., Colombia. mmercado@ins.gov.co.Grupo de Inmunovirología, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia. maria.rugeles@udea.edu.co.

Pub Type(s)

Journal Article

Language

eng spa

PubMed ID

33152198

Citation

Díaz, Francisco J., et al. "Isolation and Characterization of an Early SARS-CoV-2 Isolate From the 2020 Epidemic in Medellín, Colombia." Biomedica : Revista Del Instituto Nacional De Salud, vol. 40, no. Supl. 2, 2020, pp. 148-158.
Díaz FJ, Aguilar-Jiménez W, Flórez-Álvarez L, et al. Isolation and characterization of an early SARS-CoV-2 isolate from the 2020 epidemic in Medellín, Colombia. Biomedica. 2020;40(Supl. 2):148-158.
Díaz, F. J., Aguilar-Jiménez, W., Flórez-Álvarez, L., Valencia, G., Laiton-Donato, K., Franco-Muñoz, C., Álvarez-Díaz, D., Mercado-Reyes, M., & Rugeles, M. T. (2020). Isolation and characterization of an early SARS-CoV-2 isolate from the 2020 epidemic in Medellín, Colombia. Biomedica : Revista Del Instituto Nacional De Salud, 40(Supl. 2), 148-158. https://doi.org/10.7705/biomedica.5834
Díaz FJ, et al. Isolation and Characterization of an Early SARS-CoV-2 Isolate From the 2020 Epidemic in Medellín, Colombia. Biomedica. 2020 10 30;40(Supl. 2):148-158. PubMed PMID: 33152198.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation and characterization of an early SARS-CoV-2 isolate from the 2020 epidemic in Medellín, Colombia AU - Díaz,Francisco J, AU - Aguilar-Jiménez,Wbeimar, AU - Flórez-Álvarez,Lizdany, AU - Valencia,Gladys, AU - Laiton-Donato,Katherine, AU - Franco-Muñoz,Carlos, AU - Álvarez-Díaz,Diego, AU - Mercado-Reyes,Marcela, AU - Rugeles,María Teresa, Y1 - 2020/10/30/ PY - 2020/09/29/received PY - 2020/11/5/entrez PY - 2020/11/6/pubmed PY - 2020/11/20/medline KW - Coronavirus infections KW - viral isolation severe acute respiratory syndrome KW - SARS virus KW - high-throughput nucleotide sequencing KW - microscopy KW - electron KW - fluorescent antibody technique KW - indirect SP - 148 EP - 158 JF - Biomedica : revista del Instituto Nacional de Salud JO - Biomedica VL - 40 IS - Supl. 2 N2 - 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. SN - 2590-7379 UR - https://www.unboundmedicine.com/medline/citation/33152198/Isolation_and_characterization_of_an_early_SARS_CoV_2_isolate_from_the_2020_epidemic_in_Medellín_Colombia L2 - https://www.revistabiomedica.org/index.php/biomedica/article/view/5834 DB - PRIME DP - Unbound Medicine ER -