Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/13666
Title: Detection and sequencing of Zika virus from amniotic fluid of fetuses with microcephaly in Brazil: a case study
Authors: Calvet, Guilherme
Aguiar, Renato S.
Melo, Adriana S. O.
Sampaio, Simone A.
Filippis, Ivano de
Fabri, Allison
Araujo, Eliane S. M.
Sequeira, Patricia C. de
Mendonça, Marcos C L de
Oliveira, Louisi de
Tschoeke, Diogo A.
Schrago, Carlos G.
Thompson, Fabiano L.
Brasil, Patricia
Santos, Flavia B. dos
Nogueira, Rita M. R.
Tanuri, Amilcar
Filippis, Ana M. B. de
Affilliation: Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Doenças Febris Agudas. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Departamento de Genética. Rio de Janeiro, RJ, Brasil.
Instituto de Pesquisa Professor Joaquim Amorim Neto. Campina Grande, PB, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Nacional de Controle e Qualidade em Saúde. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Departamento de Genética. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Departamento de Genética. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Laboratório de Sistemas Avançados de Gestão de Produção. Centro de Gestão Tecnológica. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Doenças Febris Agudas. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Departamento de Genética. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivírus. Rio de Janeiro, RJ, Brasil.
Abstract: Background The incidence of microcephaly in Brazil in 2015 was 20 times higher than in previous years. Congenital microcephaly is associated with genetic factors and several causative agents. Epidemiological data suggest that microcephaly cases in Brazil might be associated with the introduction of Zika virus. We aimed to detect and sequence the Zika virus genome in amniotic fluid samples of two pregnant women in Brazil whose fetuses were diagnosed with microcephaly. Methods In this case study, amniotic fl uid samples from two pregnant women from the state of Paraíba in Brazil whose fetuses had been diagnosed with microcephaly were obtained, on the recommendation of the Brazilian health authorities, by ultrasound-guided transabdominal amniocentesis at 28 weeks’ gestation. The women had presented at 18 weeks’ and 10 weeks’ gestation, respectively, with clinical manifestations that could have been symptoms of Zika virus infection, including fever, myalgia, and rash. After the amniotic fl uid samples were centrifuged, DNA and RNA were extracted from the purifi ed virus particles before the viral genome was identified by quantitative reverse transcription PCR and viral metagenomic next-generation sequencing. Phylogenetic reconstruction and investigation of recombination events were done by comparing the Brazilian Zika virus genome with sequences from other Zika strains and from flaviviruses that occur in similar regions in Brazil. Findings We detected the Zika virus genome in the amniotic fl uid of both pregnant women. The virus was not detected in their urine or serum. Tests for dengue virus, chikungunya virus, Toxoplasma gondii, rubella virus, cytomegalovirus, herpes simplex virus, HIV, Treponema pallidum, and parvovirus B19 were all negative. After sequencing of the complete genome of the Brazilian Zika virus isolated from patient 1, phylogenetic analyses showed that the virus shares 97–100% of its genomic identity with lineages isolated during an outbreak in French Polynesia in 2013, and that in both envelope and NS5 genomic regions, it clustered with sequences from North and South America, southeast Asia, and the Pacifi c. After assessing the possibility of recombination events between the Zika virus and other flaviviruses, we ruled out the hypothesis that the Brazilian Zika virus genome is a recombinant strain with other mosquito-borne flaviviruses. Interpretation These findings strengthen the putative association between Zika virus and cases of microcephaly in neonates in Brazil. Moreover, our results suggest that the virus can cross the placental barrier. As a result, Zika virus should be considered as a potential infectious agent for human fetuses. Pathogenesis studies that confi rm the tropism of Zika virus for neuronal cells are warranted.
Keywords: Sequencing of Zika virus
Zika virus
Microcephaly
Brazil
keywords: Vírus Zika
Zika
Brasil
Microcefalia
DeCS: Zika virus
Microcefalia
Issue Date: 2016
Citation: CALVET, Guilherme; AGUIAR, Renato S.; MELO, Adriana S. O.; SAMPAIO, Simone A.; FILIPPIS, Ivano de; FABRI, Allison; ARAUJO, Eliane S. M.; SEQUEIRA, Patricia C de; MENDONÇA, Marcos C. L. de; OLIVEIRA, Louisi de; TSCHOEKE, Diogo A.; SCHRAGO, Carlos G.; THOMPSON, Fabiano L.; BRASIL, Patricia; SANTOS, Flavia B. dos; NOGUEIRA, Rita M. R.; TANURI, Amilcar; FILIPPIS, Ana M. B. de. Detection and sequencing of Zika virus from amniotic fluid of fetuses with microcephaly in Brazil: a case study. The Lancet Infection Diseases, New York, p. 1-8, 18 Feb. 2016. Disponível em: <http://www.thelancet.com/pdfs/journals/laninf/PIIS1473-3099(16)00095-5.pdf>. Acesso em: 04 abr. 2016.
Description: Artigo liberado em acesso aberto como parte do acordo para tornar público todos os dados produzidos sobre o vírus zika - Compartilhamento de dados em emergências de saúde pública - http://www.wellcome.ac.uk/News/Media-office/Press-releases/2016/WTP060169.htm
DOI: 10.1016/S1473-3099(16)00095-5
ISSN: 0140-6736
Copyright: open access
Appears in Collections:IOC - Artigos de Periódicos
INCQS - Artigos de Periódicos
INI - Artigos de Periódicos



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