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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2018.tde-13082018-165426
Document
Author
Full name
Shahab Zaki Pour
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2018
Supervisor
Committee
Zanotto, Paolo Marinho de Andrade (President)
Pierulivo, Enrique Mario Boccardo
Pinho, João Renato Rebello
Souza, Renato Pereira de
Title in Portuguese
Desenvolvimento e validação experimental de uma metodologia in house para amplificação e sequenciamento do genoma completo do Zika vírus.
Keywords in Portuguese
Amplificação de genoma viral completo
Enriquecimento prévio
Reação em cadeia da polimerase
Sanger
Sequenciamento de nova geração
Zika
Abstract in Portuguese
O zika é um arbovírus emergente. Há evidências para a relação entre o zika e a microcefalia congênita e também com a síndrome de Guillain-Barre. Várias características do vírus são importantes, como a persistência do vírus no sêmen por vários meses, transmissão sexual e evidência de transmissão pré-natal. As mães grávidas infectadas com zika podem dar à luz crianças aparentemente saudáveis que podem apresentar manifestações e complicações tardias. Existe uma clara necessidade de diagnosticar e sequenciar amostras clínicas do ZIKV que circulam na América do Sul, especificamente no Brasil. No entanto, as baixas cargas virais observadas que são observadas comumente em amostras humanas constituem um fator complicador para detecção, amplificação e sequenciamento. Neste projeto, propor projetar um fluxo de trabalho otimizado para o sequenciamento completo do genoma com base no pré-enriquecimento por PCR (reação em cadeia da polimerase) e pools de amplicons.
Title in English
Development and validation of an in-house method for whole genome amplification and sequencing of Zika virus.
Keywords in English
Enrichment
New generation sequencing
Polymerase chain reaction
Sanger
Whole genome amplification
Zika
Abstract in English
Zika is an emerging arbovirus. There is enough evidence for the relation between Zika and congenital microcephaly and also with the Guillain-Barre syndrome. Several characteristics of the virus are important, such as persistence of the virus in semen for several months, sexual transmission and evidence of prenatal transmission. Zika infected pregnant mothers may give birth to apparently healthy children that may show late manifestations and complications. There is a clear necessity of diagnosing and sequencing clinical samples of ZIKV circulating in South America, specifically in Brazil. Nevertheless, the observed low viral loads that are commonly in human samples constitute a complicating factor for detection, amplification and sequencing. In this project, we aim to design an optimized workflow for full genome sequencing based on pre-enrichment by PCR (polymerase chain reaction) and amplicon pools.
 
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Publishing Date
2018-09-03
 
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