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      Characterization of local SARS-CoV-2 isolates and pathogenicity in IFNAR−/- mice

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      Author(s)
      Hanifehnezhad, A.
      Kehribar, Ebru Şahin
      Öztop, S.
      Sheraz, Ali
      Kasırga, Serkan
      Ergünay, K.
      Önder, S.
      Yılmaz, E.
      Engin, D.
      Oğuzoğlu, T. Ç.
      Şeker, Urartu Özgür Şafak
      Yılmaz, E.
      Özkul, A.
      Date
      2020
      Source Title
      Heliyon
      Print ISSN
      2405-8440
      Publisher
      Elsevier
      Volume
      6
      Issue
      9
      Pages
      e05116
      Language
      English
      Type
      Article
      Item Usage Stats
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      12
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      Abstract
      The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) recently a global pandemic with unprecedented public health, economic and social impact. The development of effective mitigation strategies, therapeutics and vaccines relies on detailed genomic and biological characterization of the regional viruses. This study was carried out to isolate SARS-CoV-2 viruses circulating in Anatolia, and to investigate virus propagation in frequently-used cells and experimental animals. We obtained two SARS-CoV-2 viruses from nasopharngeal swabs of confirmed cases in Vero E6 cells, visualized the virions using atomic force and scanning electron microscopy and determined size distribution of the particles. Viral cytopathic effects on Vero E6 cells were initially observed at 72 h post-inoculation and reached 90% of the cells on the 5th day. The isolates displayed with similar infectivity titers, time course and infectious progeny yields. Genome sequencing revealed the viruses to be well-conserved, with less than 1% diversity compared to the prototype virus. The analysis of the viral genomes, along with the available 62 complete genomes from Anatolia, showed limited diversity (up to 0.2% on deduced amino acids) and no evidence of recombination. The most prominent sequence variation was observed on the spike protein, resulting in the substitution D614G, with a prevalence of 56.2%. The isolates produced non-fatal infection in the transgenic type I interferon knockout (IFNAR−/-) mice, with varying neutralizing antibody titers. Hyperemia, regional consolidation and subpleural air accumulation was observed on necropsy, with similar histopathological and immunohistochemistry findings in the lungs, heart, stomach, intestines, liver, spleen and kidneys. Peak viral loads were detected in the lungs, with virus RNA present in the kidneys, jejunum, liver, spleen and heart. In conclusion, we characterized two local isolates, investigated in vitro growth dynamics in Vero E6 cells and identified IFNAR−/− mice as a potential animal model for SARS-CoV-2 experiments.
      Keywords
      Bioinformatics
      Microbiology
      Virology
      Viral disease
      Viral genetics
      Epidemiology
      Respiratory system
      SARS-CoV-2
      Isolate
      Genome
      IFNAR -/- mice
      Permalink
      http://hdl.handle.net/11693/75543
      Published Version (Please cite this version)
      https://dx.doi.org/10.1016/j.heliyon.2020.e05116
      Collections
      • Institute of Materials Science and Nanotechnology (UNAM) 1930
      • Nanotechnology Research Center (NANOTAM) 1063
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