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Master's Dissertation
DOI
10.11606/D.10.2013.tde-05062013-150800
Document
Author
Full name
Bruna Rocha Passos Barbosa
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Gregori, Fábio (President)
Jerez, José Antonio
Linares, Rita de Cássia
Title in Portuguese
Caracterização e análise molecular dos genes codificadores das proteínas não estruturais 2 e 5 (NSP2 e NSP5) de rotavírus suínos
Keywords in Portuguese
Diagnóstico
Medicina Veterinária Preventiva
PCR
Rotavírus
Suíno
Abstract in Portuguese
Os rotavírus são os responsáveis pela ocorrência de diarreias em humanos e outras diversas espécies animais. Estão amplamente disseminados na suinocultura, inclusive no Brasil. As proteínas não estruturais 2 e 5 (NSP2 e NSP5) dos rotavírus estão envolvidas nas etapas de replicação viral, sendo essenciais para a formação do viroplasma, uma estrutura citoplasmática no interior da qual ocorre a morfogênese das novas partículas virais. Entretanto, são escassos os estudos sobre a diversidade genética destas proteínas em rotavírus circulantes nas criações brasileiras. Até o presente momento, a NSP2 pode ser classificada em nove genotipos (N1 ao N9) e a NSP5, 11 (H1 ao H11), sendo que em humanos foram descritos os genotipos N1, N2, N3 e H1, H2 e H3, e em suínos N1 e H1. Este estudo teve o objetivo de caracterizar as amostras circulantes de rotavírus em termos da diversidade da NSP2 e NSP5. Para isso, um total de 63 amostras fecais provenientes de criações de suínos localizadas em seis diferentes municípios do Estado de São Paulo, Brasil, foram previamente triadas mediante a técnica de nested-PCR. Destas, nove tiveram os respectivos segmentos genômicos amplificados pela reação de RT-PCR, sendo que em sete foi possível o sequenciamento nucleotídico parcial para NSP2 e, em seis, o sequenciamento total para NSP5. Todas foram caracterizadas como genotipo N1 e H1. Considerando o gene NSP2, nas amostras aqui definidas, a identidade nucleotídica variou de 100% a 86,4%, e em termos de aminoácidos, de 100% a 91,5%, enquanto que para NSP5 foi de 100% a 95,1%, e de 100% a 97,4% respectivamente para nucleotídeos e aminoácidos. Conclui-se que os genotipos das amostras circulantes na região de estudo estão em concordância com aqueles descritos na literatura para a espécie suína, e que há a hipótese de interação entre rotavirus de origem humana e animal. Estes dados são úteis para uma vigilância mais abrangente dos rotavírus circulantes e contribuem para uma melhor compreensão da patogenia, epidemiologia e prevenção da doença, inclusive no que diz respeito ao seu caráter zoonótico.
Title in English
Characterization and molecular analysis of genes coding for non-structural proteins 2 and 5 (NSP2 and NSP5) of swine rotavirus
Keywords in English
Diagnostic
PCR
Preventive Veterinary Medicine
Rotavirus
Swine
Abstract in English
Rotaviruses are responsible for the occurrence of diarrhea in humans and several other animal species. They are widespread in pig farms, including in Brazil. The non-structural proteins 2 and 5 (NSP2 and NSP5) of rotavirus are involved in viral replication and they are essential for the formation of viroplasm, a cytoplasmic structure within which occurs morphogenesis of new viral particles. However, there are very few studies on the genetic diversity of those proteins in circulating rotavirus in Brazilian swine raisings. So far, nine NSP2 genotypes have been identified (N1 to N9) and eleven for NSP5 (H1 to H11). In humans, genotypes N1, N2, N3 and H1, H2, H3 have been described, whereas in pigs, H1 and N1 have been described. This study is aimed at characterizing circulating samples of rotavirus in terms of diversity of NSP2 and NSP5. For this purpose, a total of 63 fecal samples from pig farms located in six different cities in the state of São Paulo, Brazil, were previously screened by nested-PCR technique. Of those, nine had their genomic segments amplified by RT-PCR, and in seven it was possible to obtain the partial nucleotide sequencing for NSP2, whereas in six, the total sequencing for NSP5. All were characterized as genotype H1 and N1. Considering the gene NSP2, the strains nucleotide identity, defined herein, ranged from 100% to 86.4% and in terms of amino acids, from 100% to 91.5%. Whereas for NSP5, it was from 100% to 95.1 %, and 100% to 97.4% for nucleotides and amino acids, respectively. It is concluded that the genotypes of the strains circulating in the region of study are in agreement with those reported in literature for swine, and that there is the possibility of interaction between human and animal rotaviruses. These data are useful for a broader surveillance of circulating rotaviruses and contribute to a better understanding of the pathogenesis, epidemiology and disease prevention, especially in regard to its zoonotic aspect.
 
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Publishing Date
2013-06-25
 
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