Monkeypox (Mpox) virus isolation and ultrastructural characterisation from a Brazilian human sample case.

Milene Dias Miranda, Gabriela Cardoso Caldas, Vivian Neuza Ferreira, Ortrud Monika Barth, Aline de Paula Dias da Silva, Mayara Secco Torres Silva, Beatriz Grinsztejn, Valdiléa Gonçalves Veloso, Thiago Moreno Souza, Edson Elias da Silva, Debora Ferreira Barreto-Vieira
Author Information
  1. Milene Dias Miranda: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil. ORCID
  2. Gabriela Cardoso Caldas: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil.
  3. Vivian Neuza Ferreira: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil.
  4. Ortrud Monika Barth: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil.
  5. Aline de Paula Dias da Silva: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil.
  6. Mayara Secco Torres Silva: Fundação Oswaldo Cruz-Fiocruz, Instituto Nacional de Infectologia Evandro Chagas, Rio de Janeiro, RJ, Brasil.
  7. Beatriz Grinsztejn: Fundação Oswaldo Cruz-Fiocruz, Instituto Nacional de Infectologia Evandro Chagas, Rio de Janeiro, RJ, Brasil.
  8. Valdiléa Gonçalves Veloso: Fundação Oswaldo Cruz-Fiocruz, Instituto Nacional de Infectologia Evandro Chagas, Rio de Janeiro, RJ, Brasil.
  9. Thiago Moreno Souza: Fundação Oswaldo Cruz-Fiocruz, Centro de Desenvolvimento Tecnológico em Saúde, Instituto Nacional de Ciência e Tecnologia de Gestão da Inovação em Doenças Negligenciadas, Rio de Janeiro, RJ, Brasil.
  10. Edson Elias da Silva: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Vírus Respiratórios, Exantemáticos, Enterovírus e Emergências Virais, Rio de Janeiro, RJ, Brasil.
  11. Debora Ferreira Barreto-Vieira: Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Morfologia e Morfogênese Viral, Rio de Janeiro, RJ, Brasil. ORCID

Abstract

BACKGROUND: According to the last 2023 Monkeypox (Mpox) Outbreak Global Map from the Centres for Disease Control and Prevention (CDC), more than 100 countries with no Mpox infection report cases. Brazil stands out in this group and is the second country with the highest number of cases in the last outbreak.
OBJECTIVE: To contribute to knowledge of the virus infection effects in a cellular model, which is important for diagnosis infections not yet included in a provider´s differential diagnosis and for developing viral inhibition strategies.
METHODS: We describe a virus isolation protocol for a human clinical sample from a patient from Brazil, the viral growth in a cell model through plaque forming units (PFU) assay, reverse transcriptase polymerase chain reaction (RT-PCR) and transmission electron microscopy (TEM).
FINDINGS: We follow the viral isolation in Vero cell culture from a Mpox positive clinically diagnosed sample and show the infection effects on cellular structures using a TEM.
MAIN CONCLUSIONS: Understanding the impact of viral growth on cellular structures and its replication kinetics may offer better strategies for the development of new drugs with antiviral properties.

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MeSH Term

Humans
Brazil
Mpox (monkeypox)
Biological Assay
Diagnosis, Differential
Disease Outbreaks

Word Cloud

Created with Highcharts 10.0.0MpoxviralinfectionviruscellularisolationsamplelastMonkeypoxcasesBrazileffectsmodeldiagnosisstrategieshumangrowthcellTEMstructuresBACKGROUND:According2023OutbreakGlobalMapCentresDiseaseControlPreventionCDC100countriesreportstandsgroupsecondcountryhighestnumberoutbreakOBJECTIVE:contributeknowledgeimportantinfectionsyetincludedprovider´sdifferentialdevelopinginhibitionMETHODS:describeprotocolclinicalpatientplaqueformingunitsPFUassayreversetranscriptasepolymerasechainreactionRT-PCRtransmissionelectronmicroscopyFINDINGS:followVeroculturepositiveclinicallydiagnosedshowusingMAINCONCLUSIONS:UnderstandingimpactreplicationkineticsmayofferbetterdevelopmentnewdrugsantiviralpropertiesultrastructuralcharacterisationBraziliancase

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