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An??lise do metagenoma viral de amostras de fezes humanas do Distrito FederalSantos, Rayane Nogueira dos 27 March 2015 (has links)
Submitted by Kelson Anthony de Menezes (kelson@ucb.br) on 2016-12-09T13:47:20Z
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Previous issue date: 2015-03-27 / Acute gastroenteritis, especially in developing countries, is a major cause of mortality and morbidity, which affects people of all social classes. The knowledge of etiological agent assists the choice of a treatment strategy, but also directs epidemiological studies for control measures and disease prevention, as the development of vaccines and diagnostic tests. Metagenomic approaches enable the detection of viral sequences to determine the virus population present in the stool. The objective of this study was to evaluate the viroma of human feces in 4 distinct groups, consisting of diarrheal stool samples from children, diarrheal from adults, no diarrhea from adults and immunocompromised, identifying likely pathogens involved in cases of gastroenteritis. The viral metagenomic execution method consists of the semi-purification, extraction of RNA and DNA, amplification of RNA and DNA, library construction, sequencing and analysis of the data by bioinformatics. In the analysis of the first group were detecting bacteriophages, astrovirus and torque teno virus; in the second group were identified, among others, bacteriophages, adenoviruses and torque teno virus; the third group of viruses Circoviridae family and bacteriophages; in the fourth group adenovirus, torque teno virus, gyrovirus and human papillomavirus. These results emphasize what has been identified in the literature and provides evidence that viral metagenomics has facilitated advances in the field of virology, for being a sensitive technique for the detection of viruses which can not be identified by traditional culture.Acute gastroenteritis, especially in developing countries, is a major cause of mortality and morbidity, which affects people of all social classes. The knowledge of etiological agent assists the choice of a treatment strategy, but also directs epidemiological studies for control measures and disease prevention, as the development of vaccines and diagnostic tests. Metagenomic approaches enable the detection of viral sequences to determine the virus population present in the stool. The objective of this study was to evaluate the viroma of human feces in 4 distinct groups, consisting of diarrheal stool samples from children, diarrheal from adults, no diarrhea from adults and immunocompromised, identifying likely pathogens involved in cases of gastroenteritis. The viral metagenomic execution method consists of the semi-purification, extraction of RNA and DNA, amplification of RNA and DNA, library construction, sequencing and analysis of the data by bioinformatics. In the analysis of the first group were detecting bacteriophages, astrovirus and torque teno virus; in the second group were identified, among others, bacteriophages, adenoviruses and torque teno virus; the third group of viruses Circoviridae family and bacteriophages; in the fourth group adenovirus, torque teno virus, gyrovirus and human papillomavirus. These results emphasize what has been identified in the literature and provides evidence that viral metagenomics has facilitated advances in the field of virology, for being a sensitive technique for the detection of viruses which can not be identified by traditional culture. / A gastroenterite aguda, especialmente em pa??ses em desenvolvimento, ?? uma importante causa de mortalidade e morbidade, que atinge pessoas de todas as classes sociais. O conhecimento do agente etiol??gico nestas infec????es auxilia a escolha da estrat??gia de tratamento, como tamb??m direciona estudos epidemiol??gicos para medidas de controle e preven????o de doen??a, como o desenvolvimento de vacinas e testes diagn??sticos. Abordagens metagen??micas possibilitam a detec????o de sequ??ncias virais, para determinar a popula????o viral presente nas fezes. Assim, o objetivo desse trabalho foi avaliar o viroma de fezes humanas em 4 grupos distintos, sendo composto por amostras fecais diarreicas de crian??as, diarreicas de adultos, n??o diarreicas de adultos e de imunocomprometidos, identificando prov??veis pat??genos envolvidos nos quadros de gastroenterite. O m??todo de execu????o da metagen??mica viral consistiu na semi-purifica????o, extra????o de RNA e DNA, amplifica????o do RNA e DNA, constru????o da biblioteca, sequenciamento e an??lises dos dados por bioinform??tica. Na an??lise do primeiro grupo houve detec????o de bacteri??fagos, astrov??rus e torque teno v??rus; no segundo grupo foram identificados, entre outros, bacteri??fagos, adenov??rus e torque teno v??rus; no terceiro grupo v??rus da fam??lia Circoviridae e de bacteri??fagos; no quarto grupo adenov??rus, torque teno v??rus, gyrovirus e papilomav??rus humano. Esses resultados enfatizam o que tem se identificado na literatura e fornece evid??ncia de que a metagen??mica viral tem facilitado os avan??os no campo da virologia, sendo esta, uma t??cnica sens??vel para a detec????o de v??rus que n??o podem ser identificados por cultura tradicional.
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