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PLoS Negl Trop Dis ; 13(4): e0007184, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31022183

RESUMO

The frequency of epidemics caused by Dengue viruses 1-4, Zika virus and Chikungunya viruses have been on an upward trend in recent years driven primarily by uncontrolled urbanization, mobility of human populations and geographical spread of their shared vectors, Aedes aegypti and Aedes albopictus. Infections by these viruses present with similar clinical manifestations making them challenging to diagnose; this is especially difficult in regions of the world hyperendemic for these viruses. In this study, we present a targeted-enrichment methodology to simultaneously sequence the complete viral genomes for each of these viruses directly from clinical samples. Additionally, we have also developed a customized computational tool (BaitMaker) to design these enrichment baits. This methodology is robust in its ability to capture diverse sequences and is amenable to large-scale epidemiological studies. We have applied this methodology to two large cohorts: a febrile study based in Colombo, Sri Lanka taken during the 2009-2015 dengue epidemic (n = 170) and another taken during the 2016 outbreak of Zika virus in Singapore (n = 162). Results from these studies indicate that we were able to cover an average of 97.04% ± 0.67% of the full viral genome from samples in these cohorts. We also show detection of one DENV3/ZIKV co-infected patient where we recovered full genomes for both viruses.


Assuntos
Vírus Chikungunya/genética , Vírus da Dengue/genética , Genoma Viral , Técnicas de Amplificação de Ácido Nucleico/métodos , Zika virus/genética , Linhagem Celular , Febre de Chikungunya/diagnóstico , Vírus Chikungunya/isolamento & purificação , Coinfecção/epidemiologia , Coinfecção/transmissão , Biologia Computacional , Dengue/diagnóstico , Vírus da Dengue/isolamento & purificação , Surtos de Doenças , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Singapura/epidemiologia , Sri Lanka/epidemiologia , Zika virus/isolamento & purificação , Infecção por Zika virus/diagnóstico
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