Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp.
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Caderno de Ciências Agrárias (Online) |
Texto Completo: | https://periodicos.ufmg.br/index.php/ccaufmg/article/view/2907 |
Resumo: | The genetic characterization of species is essential when you want to elect a vaccine strain or control the spread of outbreaks of a disease in a region. The 16S rRNA and pld Corynebacterium pseudotuberculosis genes play an important role as markers for bacterial genotype identification. The aim of this study was to validate an “in house” molecular methodology for genetic identification of Corynebacterium spp. isolated from clinical samples maintained in the laboratory. For this purpose, eleven isolates were reactivated in Brain Heart Infusion broth. The grown colonies identified as Gram-positive bacilli were submitted to chromosomal DNA extraction with KAPAExtract® kit according to manufacturer's recommendations. The extracted chromosomal DNA was quantified by 1.5% agarose gel electrophoresis and then subjected to duplex PCR with specific primers for the genus and species of Corynebacterium pseudotuberculosis. The amplicons generated were analyzed by 2.0% agarose gel electrophoresis. None of the isolated amplified the target genes for genus and specific species for those expected to be C. pseudotuberculosis. The positive control, C. pseudotuberculosis CIP102968 amplified fragments of 813 and 201 base pairs which corresponding to the 16S rRNA (genus specific) and pld genes (species specific) respectively. The negative control, Escherichia coli ATCC® 25922 do not amplified any target genes as expected. The results showed the need for improvement in the sampling techniques and previous microbiological identification preceding the molecular identification step and they are extremely important for precise genotype identification of strains of C. pseudotuberculosis. |
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Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp.Identificação genotípicaCorynebacterium16S rRNAFospolipase D.Produção AnimalThe genetic characterization of species is essential when you want to elect a vaccine strain or control the spread of outbreaks of a disease in a region. The 16S rRNA and pld Corynebacterium pseudotuberculosis genes play an important role as markers for bacterial genotype identification. The aim of this study was to validate an “in house” molecular methodology for genetic identification of Corynebacterium spp. isolated from clinical samples maintained in the laboratory. For this purpose, eleven isolates were reactivated in Brain Heart Infusion broth. The grown colonies identified as Gram-positive bacilli were submitted to chromosomal DNA extraction with KAPAExtract® kit according to manufacturer's recommendations. The extracted chromosomal DNA was quantified by 1.5% agarose gel electrophoresis and then subjected to duplex PCR with specific primers for the genus and species of Corynebacterium pseudotuberculosis. The amplicons generated were analyzed by 2.0% agarose gel electrophoresis. None of the isolated amplified the target genes for genus and specific species for those expected to be C. pseudotuberculosis. The positive control, C. pseudotuberculosis CIP102968 amplified fragments of 813 and 201 base pairs which corresponding to the 16S rRNA (genus specific) and pld genes (species specific) respectively. The negative control, Escherichia coli ATCC® 25922 do not amplified any target genes as expected. The results showed the need for improvement in the sampling techniques and previous microbiological identification preceding the molecular identification step and they are extremely important for precise genotype identification of strains of C. pseudotuberculosis.Universidade Federal de Minas Gerais2016-08-31info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://periodicos.ufmg.br/index.php/ccaufmg/article/view/2907Agrarian Sciences Journal; Vol. 8 No. 2 (2016); 23-29Caderno de Ciências Agrárias; v. 8 n. 2 (2016); 23-292447-62181984-6738reponame:Caderno de Ciências Agrárias (Online)instname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGporhttps://periodicos.ufmg.br/index.php/ccaufmg/article/view/2907/1758Copyright (c) 2016 Alessandra Rejane Ericsson de Oliveira Xavier, Angélica Alves de Moura Freitas, Anna Christina de almeida, Vasco Ariston de Carvalho Azevedo, Igor Viana Brandiinfo:eu-repo/semantics/openAccessEricsson de Oliveira Xavier, Alessandra Rejanede Moura Freitas, Angélica Alvesde almeida, Anna Christinade Carvalho Azevedo, Vasco AristonBrandi, Igor Viana2019-02-01T19:29:55Zoai:periodicos.ufmg.br:article/2907Revistahttps://periodicos.ufmg.br/index.php/ccaufmgPUBhttps://periodicos.ufmg.br/index.php/ccaufmg/oaiccaufmg@ica.ufmg.br2447-62181984-6738opendoar:2019-02-01T19:29:55Caderno de Ciências Agrárias (Online) - Universidade Federal de Minas Gerais (UFMG)false |
dc.title.none.fl_str_mv |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
title |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
spellingShingle |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. Ericsson de Oliveira Xavier, Alessandra Rejane Identificação genotípica Corynebacterium 16S rRNA Fospolipase D. Produção Animal |
title_short |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
title_full |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
title_fullStr |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
title_full_unstemmed |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
title_sort |
Genes 16S RNA ribossomal e pld como marcadores moleculares para identificação genotípica de amostras clínicas de Corynebacterium spp. |
author |
Ericsson de Oliveira Xavier, Alessandra Rejane |
author_facet |
Ericsson de Oliveira Xavier, Alessandra Rejane de Moura Freitas, Angélica Alves de almeida, Anna Christina de Carvalho Azevedo, Vasco Ariston Brandi, Igor Viana |
author_role |
author |
author2 |
de Moura Freitas, Angélica Alves de almeida, Anna Christina de Carvalho Azevedo, Vasco Ariston Brandi, Igor Viana |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Ericsson de Oliveira Xavier, Alessandra Rejane de Moura Freitas, Angélica Alves de almeida, Anna Christina de Carvalho Azevedo, Vasco Ariston Brandi, Igor Viana |
dc.subject.por.fl_str_mv |
Identificação genotípica Corynebacterium 16S rRNA Fospolipase D. Produção Animal |
topic |
Identificação genotípica Corynebacterium 16S rRNA Fospolipase D. Produção Animal |
description |
The genetic characterization of species is essential when you want to elect a vaccine strain or control the spread of outbreaks of a disease in a region. The 16S rRNA and pld Corynebacterium pseudotuberculosis genes play an important role as markers for bacterial genotype identification. The aim of this study was to validate an “in house” molecular methodology for genetic identification of Corynebacterium spp. isolated from clinical samples maintained in the laboratory. For this purpose, eleven isolates were reactivated in Brain Heart Infusion broth. The grown colonies identified as Gram-positive bacilli were submitted to chromosomal DNA extraction with KAPAExtract® kit according to manufacturer's recommendations. The extracted chromosomal DNA was quantified by 1.5% agarose gel electrophoresis and then subjected to duplex PCR with specific primers for the genus and species of Corynebacterium pseudotuberculosis. The amplicons generated were analyzed by 2.0% agarose gel electrophoresis. None of the isolated amplified the target genes for genus and specific species for those expected to be C. pseudotuberculosis. The positive control, C. pseudotuberculosis CIP102968 amplified fragments of 813 and 201 base pairs which corresponding to the 16S rRNA (genus specific) and pld genes (species specific) respectively. The negative control, Escherichia coli ATCC® 25922 do not amplified any target genes as expected. The results showed the need for improvement in the sampling techniques and previous microbiological identification preceding the molecular identification step and they are extremely important for precise genotype identification of strains of C. pseudotuberculosis. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-08-31 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://periodicos.ufmg.br/index.php/ccaufmg/article/view/2907 |
url |
https://periodicos.ufmg.br/index.php/ccaufmg/article/view/2907 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://periodicos.ufmg.br/index.php/ccaufmg/article/view/2907/1758 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
dc.source.none.fl_str_mv |
Agrarian Sciences Journal; Vol. 8 No. 2 (2016); 23-29 Caderno de Ciências Agrárias; v. 8 n. 2 (2016); 23-29 2447-6218 1984-6738 reponame:Caderno de Ciências Agrárias (Online) instname:Universidade Federal de Minas Gerais (UFMG) instacron:UFMG |
instname_str |
Universidade Federal de Minas Gerais (UFMG) |
instacron_str |
UFMG |
institution |
UFMG |
reponame_str |
Caderno de Ciências Agrárias (Online) |
collection |
Caderno de Ciências Agrárias (Online) |
repository.name.fl_str_mv |
Caderno de Ciências Agrárias (Online) - Universidade Federal de Minas Gerais (UFMG) |
repository.mail.fl_str_mv |
ccaufmg@ica.ufmg.br |
_version_ |
1797042442781851648 |