Desenvolvimento de metodologia high-throughput para estudo populacional do mosquito Aedes aegypti e comparação de dados de genes mitocondriais
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Data
2011-04-06
Autores
Spenassatto, Carine [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O Aedes aegypti, culicídeo de hábitos diurnos, é originário do continente africano e está globalmente distribuído pelos trópicos em associação com as populações humanas. É considerado de grande importância epidemiológica por ser o principal vetor dos quatro sorotipos do vírus da dengue e da febre amarela. Uma das primeiras detecções da presença do mosquito no Estado de São Paulo aconteceu na década de 80 na cidade de Santos. Atualmente não há disponível nenhuma vacina ou medicamento eficiente contra a dengue, assim o controle da doença está restrito ao controle do vetor. Uma das alternativas de controle e entendimento das relações vetor-patógeno-homem se baseiam no desenvolvimento de ferramentas moleculares que utilizam técnicas baseadas em PCR, as quais têm possibilitado o estudo genético das populações do Ae. aegypti. Em tais estudos, vários marcadores foram envolvidos, tais como os SNPs (Single Nucleotide Polymorphisms) e marcadores mitocondriais. Nós desenvolvemos ensaios utilizando metodologia TaqMan® para o estudo genético populacional de duas populações do mosquito Aedes aegypti de cidades portuárias do Estado de São Paulo, utilizando nove marcadores SNPs. Verificamos que esta metodologia é reprodutível, rápida, de baixo custo e eficiente para estudos em larga escala. Pela análise AMOVA encontramos uma baixa, mas significativa diferenciação genética entre as populações do estudo (FST = 0,0324; P < 0,01), e uma alta taxa de migrantes por geração (8,72 entre as populações 2007 e 5,39 entre as populações 2008), indicando fluxo gênico entre as populações. A análise implementada no software Structure 2.3.1, evidenciou a existência de três clusters baseados em semelhanças genotípicas, distribuídos em dois grupos, confirmando uma moderada estruturação populacional. Verificamos através da análise de fragmentos...
Aedes aegypti, is a diurnal mosquito, originated from the African continent and is globally distributed through the tropics in association with human populations. It is considered of great epidemiological importance for being the main vector of the four serotypes of Dengue and Yellow Fever. One of the first detections of the presence of the mosquito in the State of São Paulo happened in the 80's, in the city of Santos. Currently there is no available vaccine or effective medicine against dengue fever, and disease control is restricted to vector control. An alternative to control and understanding of vectorpathogen- man relationships are based on the development of molecular tools that use PCR-based techniques, which have enabled the genetic study of populations of Ae. aegypti. In such studies, several markers were involved, such as SNPs (Single Nucleotide Polymorphisms) and mitochondrial ones. We have developed assays using TaqMan® methodology for population genetic studies of two populations of Aedes aegypti from the port cities of São Paulo, using nine SNPs markers. We found that this methodology is reproducible, fast, inexpensive and efficient for large-scale studies. AMOVA analysis found a low but significant genetic differentiation between the studied populations (FST = 0.0324, P <0.01), and a high rate of migrants per generation (8.72 among populations in 2007 and 5.39 among populations in 2008), indicating gene flow between populations. The analysis implemented in software Structure 2.3.1, revealed the existence of three clusters based on genotypic similarities, divided into two groups, confirming a moderate population structure. We verified through the analysis of the mitochondrial gene fragments NADH dehydrogenase subunit 4 (ND4) a high genetic differentiation between the two study populations (FST = 0.18034, P <0.01), and a rate of migrants per generation considered high... (Complete abstract click electronic access below)
Aedes aegypti, is a diurnal mosquito, originated from the African continent and is globally distributed through the tropics in association with human populations. It is considered of great epidemiological importance for being the main vector of the four serotypes of Dengue and Yellow Fever. One of the first detections of the presence of the mosquito in the State of São Paulo happened in the 80's, in the city of Santos. Currently there is no available vaccine or effective medicine against dengue fever, and disease control is restricted to vector control. An alternative to control and understanding of vectorpathogen- man relationships are based on the development of molecular tools that use PCR-based techniques, which have enabled the genetic study of populations of Ae. aegypti. In such studies, several markers were involved, such as SNPs (Single Nucleotide Polymorphisms) and mitochondrial ones. We have developed assays using TaqMan® methodology for population genetic studies of two populations of Aedes aegypti from the port cities of São Paulo, using nine SNPs markers. We found that this methodology is reproducible, fast, inexpensive and efficient for large-scale studies. AMOVA analysis found a low but significant genetic differentiation between the studied populations (FST = 0.0324, P <0.01), and a high rate of migrants per generation (8.72 among populations in 2007 and 5.39 among populations in 2008), indicating gene flow between populations. The analysis implemented in software Structure 2.3.1, revealed the existence of three clusters based on genotypic similarities, divided into two groups, confirming a moderate population structure. We verified through the analysis of the mitochondrial gene fragments NADH dehydrogenase subunit 4 (ND4) a high genetic differentiation between the two study populations (FST = 0.18034, P <0.01), and a rate of migrants per generation considered high... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Aedes aegypti - Genética, Population genetics
Como citar
SPENASSATTO, Carine. Desenvolvimento de metodologia high-throughput para estudo populacional do mosquito Aedes aegypti e comparação de dados de genes mitocondriais. 2011. 96 f. Dissertação (mestrado) - Universidade Estadual Paulista, Instituto de Biociências de Botucatu, 2011.