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1.
PLoS Negl Trop Dis ; 10(6): e0004816, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27341420

RESUMO

BACKGROUND: Zika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil. METHODOLOGY/PRINCIPAL FINDINGS: Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak. CONCLUSIONS/SIGNIFICANCE: The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution of alternative non-vectorial ZIKV transmission routes, needs further investigation.


Assuntos
Saliva/virologia , Infecção por Zika virus/diagnóstico , Infecção por Zika virus/urina , Zika virus/isolamento & purificação , Adulto , Idoso , Brasil/epidemiologia , Feminino , Genoma Viral , Humanos , Pessoa de Meia-Idade , Filogenia , Gravidez , RNA Viral/classificação , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto Jovem , Infecção por Zika virus/epidemiologia
2.
Virology ; 452-453: 202-11, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24606697

RESUMO

We have previously designed a method to construct viable recombinant Yellow Fever (YF) 17D viruses expressing heterologous polypeptides including part of the Simian Immunodeficiency Virus (SIV) Gag protein. However, the expressed region, encompassing amino acid residues from 45 to 269, was genetically unstable. In this study, we improved the genetic stability of this recombinant YF 17D virus by introducing mutations in the IRES element localized at the 5' end of the SIV gag gene. The new stable recombinant virus elicited adaptive immune responses similar to those induced by the original recombinant virus. It is, therefore, possible to increase recombinant stability by removing functional motifs from the insert that may have deleterious effects on recombinant YF viral fitness.


Assuntos
Vacinas contra a AIDS/genética , Produtos do Gene gag/genética , Infecções por HIV/virologia , Vírus da Imunodeficiência Símia/genética , Vírus da Febre Amarela/genética , Vacinas contra a AIDS/química , Vacinas contra a AIDS/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Citocinas/imunologia , Feminino , Produtos do Gene gag/imunologia , Vetores Genéticos/genética , Vetores Genéticos/imunologia , Infecções por HIV/imunologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Vírus da Imunodeficiência Símia/química , Vírus da Imunodeficiência Símia/imunologia , Vírus da Febre Amarela/imunologia
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