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haddad_sr_me_botfmvz.pdf: 305893 bytes, checksum: 02ff7e2de8b8a9ad8c10e7f22f207754 (MD5) === Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) === Universidade Estadual Paulista (UNESP) === O presente trabalho objetivou comparar, utilizando dados de um experimento de Microarray com um delineamento simples, os resultados de diferentes testes estatísticos a fim de verificar suas características na detecção de diferenças no nível de expressão dos genes. Os dados foram provenientes da South Dakota State University-EUA, do Department of Biology and Microbiology, Department of Animal Science, onde toda a parte experimental foi realizada. O material biológico envolveu quatro aves infectadas e quatro não infectadas com o vírus de bronquite infecciosa (IBV). O RNA utilizado foi extraído da camada epitelial da traquéia de animais controle e infectados com o vírus da IBV e, após a transcrição reversa foi marcado com os corantes fluorescentes (Cy3 e Cy5) e hibridizados com o microarray 13k cDNA de aves (FHCRC, Seattle, WA). A análise de dados dos resultados do experimento de microarray englobou dois estágios, sendo o primeiro denominado de Normalização, em que os dados foram pré-processados utilizando o procedimento Loess. A seguir foram realizadas as análises estatísticas propriamente ditas com testes de significância. Utilizou-se um modelo simples de ANOVA e aplicaram-se diferentes metodologias de análise. A análise das imagens revelou que dos 16192 spots em cada slide, apenas 10.926 puderam ser lidos sem defeitos no primeiro slide, 11.633 no segundo slide, 12577 no terceiro e 13.154 no quarto slide. A grande maioria dos spots em branco e controles negativos apresentou defeitos que determinaram sua eliminação. Um total de 13.597 spots foi lido no conjunto dos quatro slides, mas apenas 9.853 spots estavam representados em todos os slides. Concluiu-se que os experimentos de microarray, por tratarem de um conjunto muito grande de observações a serem analisados requerem análises estatísticas específicas. O método de Cui et al. (2005) reduziu... === The aim of this research was to compare, using real data of an experiment of Microarray with a simple design, the results of different statistical tests in order to verify their characteristics in the detection of differences in the level of expression of the genes. The data were coming of South Dakota State University-EUA, of the Department of Biology and Microbiology, Department Animal of Science, where the whole experimental part was accomplished. The biological material involved four infected animals and four no infected with the virus of infectious bronchitis (IBV). Used RNA was extracted of the layer epitelial of the windpipe of animals control and infected with the virus of IBV and, after the reverse transcription it was marked with the fluorescent colors (Cy3 and Cy5) and hybridization with the microarray 13k cDNA of birds (FHCRC, Seattle, WA). The analysis of data of the results of the microarray experiment included two apprenticeships, being the first denominated of Normalization, in that the data were pre-processed using the procedure Loess. To follow the statistical analyses they were accomplished properly said through real data with significant tests. A simple model of ANOVA was used and different analysis methodologies were applied. The analysis of the images revealed that of the 16192 spots in each slide, only 10.926 could be read without defects in the first slide, 11.633 in the second slide, 12577 in the third slide and 13.154 in the fourth slide. The great majority of the spots in white and negative controls presented defects that determined it elimination. A total of 13.597 spots was read in the group of the four slides, but only 9.853 spots were represented in all of the slides. It was ended that the microarray experiments, for they treat of a very big group of observations to be analyzed request specific statistical analyses. The method of Cui et al. (2005) it reduced... (Complete abstract click electronic access below)
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