Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro

Detalhes bibliográficos
Autor(a) principal: Bona, Mariana Duarte
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/32839
Resumo: Shigella and Escherichia coli Enteroinvasive (EIEC) constitute the pathotype Shigella/EIEC that cause Shigellosis. This infection is an important world public health problem, mainly in the children in development countries, because its clinic severity, lack of vaccine and high rate of antimicrobial resistance. This study aimed to develop primers for virulence-related genes and to evaluate clinical symptoms of Shigella/EIEC intestinal infection of children from case-control study on brazilian Semiarid. It was carried out a case-control study in six different cities of semiarid with 1200 children with 2 to 36 age months, distributed in the case group, when they had diarrheal episodes in the last 24 hours, and in the control group, when they had not diarrheal episodes. Fecal samples of each children were colected and its DNA was extracted for pathogens diagnosis by Luminex Bio-Plex® 200 (Bio-Rad, Califórnia, EUA), which 148 (12.33%) samples were obtained with positive DNA for Shigella/EIEC. From these 148 samples, fecal DNA was reextracted and the diagnosis was made by conventional PCR. The positive DNA for the pathotype was used for the detection of virulence genes. The clinic, demografics and socioeconomics dates were obtained by questionnaire answered by children parents or responsables. The dates of co-infectants enteropathogens with this pathotype were obtained from Luminex detections results. It was detected the pathotype in 7% (42/600) of case group children and in 3% (18/600) of control group, with association of pathotype detection and presence of diarrhea (p = 0.0021, OR = 2.43, IC95% 1.384 to 4.279). It was observed that 91.6% (55/60) of these pathotype positive samples had co-infections with least one other enteropathogen. There was association of co-infection with Salmonella spp. with abdominal pain presence (p = 0.0262; OR = 5.75; IC95% = 1.362 to 20.73). It was analyzed 28 virulence gens, of which the most prevalent is iucB gen (90%, 54/60). Some of these gens was associated with diarrhea cases: the gene sen (p = 0.0267; OR = 4.25; IC95% = 1.27 to 14.15), which encodes the ShET1 enterotoxin; genes ipgB2 (p = 0.0267; OR = 4.25; IC95% = 1.27 to 14.15) and ipgB1 (p= 0.0354; OR = 3.66; IC95% = 1.12 to 11,9), which encodes the ruffling of membrane of host cells; and the gene ospF (p= 0,0354; OR = 3,66; IC95% = 1,12-11,9), which encodes the suppression of host immune response. About clinical manifestations addicted to diarrhea, it was observed that the infected and symptomatic children had mild symptoms. The results demonstrated high frequence of Shigella/EIEC co-infections with least one other enteropathogen. The infection with Salmonella spp was associated with abdominal pain (p = 0.0262; OR = 5.75; 95% CI = 1.36 to 20.73. The results showed the association of diarrhea presence on these children with pathotype detection. The evaluation of presence of sen, ipgB1, ipgB2 e ospF genes suggest the association of factors enconding by these genes with impact in diarrhea. Further, the presence of factor related with pathotype intra and intercellular spreading was associated with fever in children. In addition, it was observed high frequence of pathotype copathogens and abdominal pain presence associated with presence of co-infection with Salmonella spp.
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spelling Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiroInfection with enteroinvasive shigella / escherichia coli, virulence and copatogen genes in Brazilian semen-controlled, semi-arid childrenSinais e SintomasFatores de VirulênciaInfecçõesShigellaShigella and Escherichia coli Enteroinvasive (EIEC) constitute the pathotype Shigella/EIEC that cause Shigellosis. This infection is an important world public health problem, mainly in the children in development countries, because its clinic severity, lack of vaccine and high rate of antimicrobial resistance. This study aimed to develop primers for virulence-related genes and to evaluate clinical symptoms of Shigella/EIEC intestinal infection of children from case-control study on brazilian Semiarid. It was carried out a case-control study in six different cities of semiarid with 1200 children with 2 to 36 age months, distributed in the case group, when they had diarrheal episodes in the last 24 hours, and in the control group, when they had not diarrheal episodes. Fecal samples of each children were colected and its DNA was extracted for pathogens diagnosis by Luminex Bio-Plex® 200 (Bio-Rad, Califórnia, EUA), which 148 (12.33%) samples were obtained with positive DNA for Shigella/EIEC. From these 148 samples, fecal DNA was reextracted and the diagnosis was made by conventional PCR. The positive DNA for the pathotype was used for the detection of virulence genes. The clinic, demografics and socioeconomics dates were obtained by questionnaire answered by children parents or responsables. The dates of co-infectants enteropathogens with this pathotype were obtained from Luminex detections results. It was detected the pathotype in 7% (42/600) of case group children and in 3% (18/600) of control group, with association of pathotype detection and presence of diarrhea (p = 0.0021, OR = 2.43, IC95% 1.384 to 4.279). It was observed that 91.6% (55/60) of these pathotype positive samples had co-infections with least one other enteropathogen. There was association of co-infection with Salmonella spp. with abdominal pain presence (p = 0.0262; OR = 5.75; IC95% = 1.362 to 20.73). It was analyzed 28 virulence gens, of which the most prevalent is iucB gen (90%, 54/60). Some of these gens was associated with diarrhea cases: the gene sen (p = 0.0267; OR = 4.25; IC95% = 1.27 to 14.15), which encodes the ShET1 enterotoxin; genes ipgB2 (p = 0.0267; OR = 4.25; IC95% = 1.27 to 14.15) and ipgB1 (p= 0.0354; OR = 3.66; IC95% = 1.12 to 11,9), which encodes the ruffling of membrane of host cells; and the gene ospF (p= 0,0354; OR = 3,66; IC95% = 1,12-11,9), which encodes the suppression of host immune response. About clinical manifestations addicted to diarrhea, it was observed that the infected and symptomatic children had mild symptoms. The results demonstrated high frequence of Shigella/EIEC co-infections with least one other enteropathogen. The infection with Salmonella spp was associated with abdominal pain (p = 0.0262; OR = 5.75; 95% CI = 1.36 to 20.73. The results showed the association of diarrhea presence on these children with pathotype detection. The evaluation of presence of sen, ipgB1, ipgB2 e ospF genes suggest the association of factors enconding by these genes with impact in diarrhea. Further, the presence of factor related with pathotype intra and intercellular spreading was associated with fever in children. In addition, it was observed high frequence of pathotype copathogens and abdominal pain presence associated with presence of co-infection with Salmonella spp.A Shigella spp. e a Escherichia coli Enteroinvasiva (EIEC) constituem o patotipo Shigella/EIEC e causam a Shigelose. Esta infecção configura-se em um importante problema de saúde pública mundial, principalmente na população infantil de países em desenvolvimento, devido a sua alta severidade clínica, inexistência de vacinas e elevada taxa de resistência antimicrobiana. Este estudo teve como objetivo desenvolver primers para detecção de genes codificadores de virulência e avaliar a clínica relacionada à infecção intestinal pelo patotipo Shigella/EIEC detectado em amostras fecais de crianças de estudo caso-controle no Semiárido Brasileiro. Foi realizado um estudo caso-controle em seis diferentes cidades no Semiárido com 1200 crianças de 2 a 36 meses de idade distribuídas no grupo caso, quando apresentavam episódios de diarréia nas 24 horas anteriores a coleta, e no grupo controle, quando não apresentavam diarreia. As amostras fecais de cada criança foram coletadas e extraído seu DNA fecal para diagnóstico de patógenos utilizando a plataforma Luminex Bio-Plex® 200 (Bio-Rad, Califórnia, EUA), onde obteve-se 148 (12,33%) amostras de fezes com DNA positivo para Shigella/EIEC. A partir destas 148 amostras, rextraíu-se o DNA fecal e foi feito o diagnóstico por PCR convecional. O DNA positivo para o patótipo foi utilizado para a pesquisa de genes de virulência. Os dados clínicos, populacionais e socioeconômicos das crianças foram obtidos por meio do questionário respondido pelos pais ou responsáveis das crianças. Os dados dos enteropatógenos co-infectantes com o patotipo foram obtidos de resultados da detecção pelo Luminex. Detectou-se o patotipo em 60% (42/70) das crianças do grupo caso e em 23% (18/78) do grupo controle, sendo associado à presença de diarreia (p = 0,0021, OR = 2,43, IC95% 1,384 a 4,279). Verificou-se que 91,6% (55/60) destas amostras positivas para o patotipo apresentavam co-infecções com pelo menos um outro enteropatógeno. Houve associação de co-infecção com Salmonella spp. e presença de dor abdominal (p = 0,0262; OR = 5,75; IC95% = 1,362 a 20,73). Foram pesquisados ao todo 28 genes de virulência, dos quais o mais prevalente foi o gene iucB (90%, 54/60) que codifica a aquisição de ferro pelos patógenos. Alguns genes detectados no estudo foram associados aos casos de diarreia: o gene sen (p = 0,0267; OR = 4,25; IC95% = 1,27 a 14,15), que codifica o fator da enterotoxina ShET1; os genes ipgB2 (p = 0,0267; OR = 4,25; IC95% = 1,27 a 14,15) e ipgB1 (p= 0,0354; OR = 3,66; IC95% = 1,12 a 11,9), que codificam o remodelamento da membrana das células do hospedeiro; e o gene ospF (p= 0,0354; OR = 3,66; IC95% = 1,12 a 11,9), que codifica a supressão da reposta pró-inflamatória do hospedeiro. Em relação as manifestações clínicas adicionais a diarreia, observou-se que as crianças do grupo caso apresentaram baixa prevalência de sintomas. Na análise do agrupamento de genes de virulência pesquisados neste estudo com dados dos sintomas clínicos, obteve-se associação de febre com presença do gene virA que codifica a disseminação intra e intercelular do patógeno (p = 0,0005; OR = 33; IC95% = 1,79 a 608). Os resultados demonstraram a associação de infecção entérica diarreica nessas crianças com a detecção de Shigella/EIEC. A avaliação da presença dos genes sen, ipgB1, ipgB2 e ospF sugerem a associação dos fatores codificados por estes genes impactando na diarreia. Além disso, a presença do gene que codifica a disseminação celular de Shigella/EIEC foi associado à febre nas crianças. Os resultados também demonstraram alta frequência de co-infecções com Shigella/EIEC e a associação desta com Salmonella spp. foi correlacionado com dor abdominal.Lima, Aldo Ângelo MoreiraBona, Mariana Duarte2018-06-13T15:10:19Z2018-06-13T15:10:19Z2018-05-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfBONA, M. D. Infecção por shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso controle no semiárido brasileiro. 2018. 114 f. Dissertação (Mestrado em Ciências Médicas) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2018.http://www.repositorio.ufc.br/handle/riufc/32839porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2019-01-16T12:06:55Zoai:repositorio.ufc.br:riufc/32839Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T19:00:27.620299Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
Infection with enteroinvasive shigella / escherichia coli, virulence and copatogen genes in Brazilian semen-controlled, semi-arid children
title Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
spellingShingle Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
Bona, Mariana Duarte
Sinais e Sintomas
Fatores de Virulência
Infecções
Shigella
title_short Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
title_full Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
title_fullStr Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
title_full_unstemmed Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
title_sort Infecção por Shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso-controle no semiárido brasileiro
author Bona, Mariana Duarte
author_facet Bona, Mariana Duarte
author_role author
dc.contributor.none.fl_str_mv Lima, Aldo Ângelo Moreira
dc.contributor.author.fl_str_mv Bona, Mariana Duarte
dc.subject.por.fl_str_mv Sinais e Sintomas
Fatores de Virulência
Infecções
Shigella
topic Sinais e Sintomas
Fatores de Virulência
Infecções
Shigella
description Shigella and Escherichia coli Enteroinvasive (EIEC) constitute the pathotype Shigella/EIEC that cause Shigellosis. This infection is an important world public health problem, mainly in the children in development countries, because its clinic severity, lack of vaccine and high rate of antimicrobial resistance. This study aimed to develop primers for virulence-related genes and to evaluate clinical symptoms of Shigella/EIEC intestinal infection of children from case-control study on brazilian Semiarid. It was carried out a case-control study in six different cities of semiarid with 1200 children with 2 to 36 age months, distributed in the case group, when they had diarrheal episodes in the last 24 hours, and in the control group, when they had not diarrheal episodes. Fecal samples of each children were colected and its DNA was extracted for pathogens diagnosis by Luminex Bio-Plex® 200 (Bio-Rad, Califórnia, EUA), which 148 (12.33%) samples were obtained with positive DNA for Shigella/EIEC. From these 148 samples, fecal DNA was reextracted and the diagnosis was made by conventional PCR. The positive DNA for the pathotype was used for the detection of virulence genes. The clinic, demografics and socioeconomics dates were obtained by questionnaire answered by children parents or responsables. The dates of co-infectants enteropathogens with this pathotype were obtained from Luminex detections results. It was detected the pathotype in 7% (42/600) of case group children and in 3% (18/600) of control group, with association of pathotype detection and presence of diarrhea (p = 0.0021, OR = 2.43, IC95% 1.384 to 4.279). It was observed that 91.6% (55/60) of these pathotype positive samples had co-infections with least one other enteropathogen. There was association of co-infection with Salmonella spp. with abdominal pain presence (p = 0.0262; OR = 5.75; IC95% = 1.362 to 20.73). It was analyzed 28 virulence gens, of which the most prevalent is iucB gen (90%, 54/60). Some of these gens was associated with diarrhea cases: the gene sen (p = 0.0267; OR = 4.25; IC95% = 1.27 to 14.15), which encodes the ShET1 enterotoxin; genes ipgB2 (p = 0.0267; OR = 4.25; IC95% = 1.27 to 14.15) and ipgB1 (p= 0.0354; OR = 3.66; IC95% = 1.12 to 11,9), which encodes the ruffling of membrane of host cells; and the gene ospF (p= 0,0354; OR = 3,66; IC95% = 1,12-11,9), which encodes the suppression of host immune response. About clinical manifestations addicted to diarrhea, it was observed that the infected and symptomatic children had mild symptoms. The results demonstrated high frequence of Shigella/EIEC co-infections with least one other enteropathogen. The infection with Salmonella spp was associated with abdominal pain (p = 0.0262; OR = 5.75; 95% CI = 1.36 to 20.73. The results showed the association of diarrhea presence on these children with pathotype detection. The evaluation of presence of sen, ipgB1, ipgB2 e ospF genes suggest the association of factors enconding by these genes with impact in diarrhea. Further, the presence of factor related with pathotype intra and intercellular spreading was associated with fever in children. In addition, it was observed high frequence of pathotype copathogens and abdominal pain presence associated with presence of co-infection with Salmonella spp.
publishDate 2018
dc.date.none.fl_str_mv 2018-06-13T15:10:19Z
2018-06-13T15:10:19Z
2018-05-14
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv BONA, M. D. Infecção por shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso controle no semiárido brasileiro. 2018. 114 f. Dissertação (Mestrado em Ciências Médicas) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2018.
http://www.repositorio.ufc.br/handle/riufc/32839
identifier_str_mv BONA, M. D. Infecção por shigella/escherichia coli enteroinvasiva, genes de virulência e copatógenos em crianças de estudo caso controle no semiárido brasileiro. 2018. 114 f. Dissertação (Mestrado em Ciências Médicas) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2018.
url http://www.repositorio.ufc.br/handle/riufc/32839
dc.language.iso.fl_str_mv por
language por
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dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
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reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
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