Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFPA |
Texto Completo: | https://repositorio.ufpa.br/jspui/handle/2011/15726 |
Resumo: | Background: Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes abscesses and ulcerative lymphangitis in equines. The aim of this study was to develop a quadruplex PCR assay that would allow simultaneous detection and biovar-typing of C. pseudotuberculosis. Methods: In the present study, genomes of C. pseudotuberculosis strains were used to identify the genes involved in the nitrate reduction pathway to improve a species identification three-primer multiplex PCR assay. The nitrate reductase gene (narG) was included in the PCR assay along with the 16S, rpoB and pld genes to enhance the diagnosis of the multiplex PCR at biovar level. Results: A novel quadruplex PCR assay for C. pseudotuberculosis species and biovar identification was developed. The results of the quadruplex PCR of 348 strains, 346 previously well-characterized clinical isolates of C. pseudotuberculosis from different hosts (goats, sheep, horse, cattle, buffalo, llamas and humans), the vaccine strain 1002 and the type strain ATCC 19410T , were compared to the results of nitrate reductase identification by biochemical test. The McNemar’s Chi-squared test used to compare the two methods used for C. pseudotuberculosis biovar identification showed no significant difference (P = 0.75) [95% CI for odds ratio (0.16–6.14)] between the quadruplex PCR and the nitrate biochemical test. Concordant results were observed for 97.13% (338 / 348) of the tested strains and the kappa value was 0.94 [95% CI (0.90–0.98)]. Conclusions: The ability of the quadruplex assay to discriminate between C. pseudotuberculosis biovar Ovis and Equi strains enhances its usefulness in the clinical microbiology laboratory. |
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2023-06-21T17:42:35Z2023-06-21T17:42:35Z2017ALMEIDA, Sintia et al. Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi. BMC Genomics, online, v. 13, n. 390, p. 1-8, 2017. DOI: https://doi.org/10.1186/s12917-017-1210-5. Disponível em: https://repositorio.ufpa.br/jspui/handle/2011/15726. Acesso em:.1471-2164https://repositorio.ufpa.br/jspui/handle/2011/1572610.1186/s12917-017-1210-5Background: Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes abscesses and ulcerative lymphangitis in equines. The aim of this study was to develop a quadruplex PCR assay that would allow simultaneous detection and biovar-typing of C. pseudotuberculosis. Methods: In the present study, genomes of C. pseudotuberculosis strains were used to identify the genes involved in the nitrate reduction pathway to improve a species identification three-primer multiplex PCR assay. The nitrate reductase gene (narG) was included in the PCR assay along with the 16S, rpoB and pld genes to enhance the diagnosis of the multiplex PCR at biovar level. Results: A novel quadruplex PCR assay for C. pseudotuberculosis species and biovar identification was developed. The results of the quadruplex PCR of 348 strains, 346 previously well-characterized clinical isolates of C. pseudotuberculosis from different hosts (goats, sheep, horse, cattle, buffalo, llamas and humans), the vaccine strain 1002 and the type strain ATCC 19410T , were compared to the results of nitrate reductase identification by biochemical test. The McNemar’s Chi-squared test used to compare the two methods used for C. pseudotuberculosis biovar identification showed no significant difference (P = 0.75) [95% CI for odds ratio (0.16–6.14)] between the quadruplex PCR and the nitrate biochemical test. Concordant results were observed for 97.13% (338 / 348) of the tested strains and the kappa value was 0.94 [95% CI (0.90–0.98)]. Conclusions: The ability of the quadruplex assay to discriminate between C. pseudotuberculosis biovar Ovis and Equi strains enhances its usefulness in the clinical microbiology laboratory.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisALVES, J. T. C.; CARNEIRO, A. R. Universidade Federal do ParáengBioMed Central LtdReino UnidoBMC GenomicsAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccesshttps://bmcvetres.biomedcentral.com/articles/10.1186/s12917-017-1210-5#Ack1reponame:Repositório Institucional da UFPAinstname:Universidade Federal do Pará (UFPA)instacron:UFPACaseous lymphadenitisDiagnosisNitrate reductaseHorseSheepGoatsQuadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equiinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1329018http://lattes.cnpq.br/6389878370640685http://lattes.cnpq.br/4646959280000344http://lattes.cnpq.br/9717412363973376http://lattes.cnpq.br/2484200467965399http://lattes.cnpq.br/2126288732219018http://lattes.cnpq.br/5069850397034946http://lattes.cnpq.br/7533716053525477http://lattes.cnpq.br/7009849807650076http://lattes.cnpq.brhttp://lattes.cnpq.br/9326996169562214http://lattes.cnpq.br/4673198445813304http://lattes.cnpq.br/3340492777093358http://lattes.cnpq.br/1020477751003832ALMEIDA, Sintia Silva deDORNELES, Elaine Maria SelesDINIZ, Carlos Augusto AlmeidaABREU, Vinicius Augusto Carvalho deSOUSA, Cassiana Severiano deALVES, Jorianne Thyeska CastroCARNEIRO, Adriana RibeiroVILAS BOAS, Priscilla Carolinne BaganoSPIER, SharonBARH, DebmalyaLAGE, Andrey PereiraFIGUEIREDO, Henrique Cesar PereiraAZEVEDO, Vasco Ariston de Carvalhohttps://orcid.org/0000-0003-0270-9059https://orcid.org/0000-0002-2753-1296https://orcid.orghttps://orcid.orghttps://orcid.orghttps://orcid.orghttps://orcid.org/0000-0003-1522-9871https://orcid.orghttps://orcid.orghttps://orcid.orghttps://orcid.orghttps://orcid.orghttps://orcid.org/0000-0002-4775-2280ORIGINALArticle_QuadruplexPcrAssay.pdfArticle_QuadruplexPcrAssay.pdfapplication/pdf623426http://repositorio.ufpa.br/oai/bitstream/2011/15726/1/Article_QuadruplexPcrAssay.pdf089d44f5171f321f374ca94dc11d9a89MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.pt_BR.fl_str_mv |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
title |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
spellingShingle |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi ALMEIDA, Sintia Silva de Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats |
title_short |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
title_full |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
title_fullStr |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
title_full_unstemmed |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
title_sort |
Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi |
author |
ALMEIDA, Sintia Silva de |
author_facet |
ALMEIDA, Sintia Silva de DORNELES, Elaine Maria Seles DINIZ, Carlos Augusto Almeida ABREU, Vinicius Augusto Carvalho de SOUSA, Cassiana Severiano de ALVES, Jorianne Thyeska Castro CARNEIRO, Adriana Ribeiro VILAS BOAS, Priscilla Carolinne Bagano SPIER, Sharon BARH, Debmalya LAGE, Andrey Pereira FIGUEIREDO, Henrique Cesar Pereira AZEVEDO, Vasco Ariston de Carvalho https://orcid.org/0000-0003-0270-9059 https://orcid.org/0000-0002-2753-1296 https://orcid.org https://orcid.org/0000-0003-1522-9871 https://orcid.org/0000-0002-4775-2280 |
author_role |
author |
author2 |
DORNELES, Elaine Maria Seles DINIZ, Carlos Augusto Almeida ABREU, Vinicius Augusto Carvalho de SOUSA, Cassiana Severiano de ALVES, Jorianne Thyeska Castro CARNEIRO, Adriana Ribeiro VILAS BOAS, Priscilla Carolinne Bagano SPIER, Sharon BARH, Debmalya LAGE, Andrey Pereira FIGUEIREDO, Henrique Cesar Pereira AZEVEDO, Vasco Ariston de Carvalho https://orcid.org/0000-0003-0270-9059 https://orcid.org/0000-0002-2753-1296 https://orcid.org https://orcid.org/0000-0003-1522-9871 https://orcid.org/0000-0002-4775-2280 |
author2_role |
author author author author author author author author author author author author author author author author author |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/6389878370640685 http://lattes.cnpq.br/4646959280000344 http://lattes.cnpq.br/9717412363973376 http://lattes.cnpq.br/2484200467965399 http://lattes.cnpq.br/2126288732219018 http://lattes.cnpq.br/5069850397034946 http://lattes.cnpq.br/7533716053525477 http://lattes.cnpq.br/7009849807650076 http://lattes.cnpq.br http://lattes.cnpq.br/9326996169562214 http://lattes.cnpq.br/4673198445813304 http://lattes.cnpq.br/3340492777093358 http://lattes.cnpq.br/1020477751003832 |
dc.contributor.author.fl_str_mv |
ALMEIDA, Sintia Silva de DORNELES, Elaine Maria Seles DINIZ, Carlos Augusto Almeida ABREU, Vinicius Augusto Carvalho de SOUSA, Cassiana Severiano de ALVES, Jorianne Thyeska Castro CARNEIRO, Adriana Ribeiro VILAS BOAS, Priscilla Carolinne Bagano SPIER, Sharon BARH, Debmalya LAGE, Andrey Pereira FIGUEIREDO, Henrique Cesar Pereira AZEVEDO, Vasco Ariston de Carvalho https://orcid.org/0000-0003-0270-9059 https://orcid.org/0000-0002-2753-1296 https://orcid.org https://orcid.org https://orcid.org https://orcid.org https://orcid.org/0000-0003-1522-9871 https://orcid.org https://orcid.org https://orcid.org https://orcid.org https://orcid.org https://orcid.org/0000-0002-4775-2280 |
dc.subject.eng.fl_str_mv |
Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats |
topic |
Caseous lymphadenitis Diagnosis Nitrate reductase Horse Sheep Goats |
description |
Background: Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes abscesses and ulcerative lymphangitis in equines. The aim of this study was to develop a quadruplex PCR assay that would allow simultaneous detection and biovar-typing of C. pseudotuberculosis. Methods: In the present study, genomes of C. pseudotuberculosis strains were used to identify the genes involved in the nitrate reduction pathway to improve a species identification three-primer multiplex PCR assay. The nitrate reductase gene (narG) was included in the PCR assay along with the 16S, rpoB and pld genes to enhance the diagnosis of the multiplex PCR at biovar level. Results: A novel quadruplex PCR assay for C. pseudotuberculosis species and biovar identification was developed. The results of the quadruplex PCR of 348 strains, 346 previously well-characterized clinical isolates of C. pseudotuberculosis from different hosts (goats, sheep, horse, cattle, buffalo, llamas and humans), the vaccine strain 1002 and the type strain ATCC 19410T , were compared to the results of nitrate reductase identification by biochemical test. The McNemar’s Chi-squared test used to compare the two methods used for C. pseudotuberculosis biovar identification showed no significant difference (P = 0.75) [95% CI for odds ratio (0.16–6.14)] between the quadruplex PCR and the nitrate biochemical test. Concordant results were observed for 97.13% (338 / 348) of the tested strains and the kappa value was 0.94 [95% CI (0.90–0.98)]. Conclusions: The ability of the quadruplex assay to discriminate between C. pseudotuberculosis biovar Ovis and Equi strains enhances its usefulness in the clinical microbiology laboratory. |
publishDate |
2017 |
dc.date.issued.fl_str_mv |
2017 |
dc.date.accessioned.fl_str_mv |
2023-06-21T17:42:35Z |
dc.date.available.fl_str_mv |
2023-06-21T17:42:35Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
ALMEIDA, Sintia et al. Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi. BMC Genomics, online, v. 13, n. 390, p. 1-8, 2017. DOI: https://doi.org/10.1186/s12917-017-1210-5. Disponível em: https://repositorio.ufpa.br/jspui/handle/2011/15726. Acesso em:. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufpa.br/jspui/handle/2011/15726 |
dc.identifier.issn.pt_BR.fl_str_mv |
1471-2164 |
dc.identifier.doi.pt_BR.fl_str_mv |
10.1186/s12917-017-1210-5 |
identifier_str_mv |
ALMEIDA, Sintia et al. Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi. BMC Genomics, online, v. 13, n. 390, p. 1-8, 2017. DOI: https://doi.org/10.1186/s12917-017-1210-5. Disponível em: https://repositorio.ufpa.br/jspui/handle/2011/15726. Acesso em:. 1471-2164 10.1186/s12917-017-1210-5 |
url |
https://repositorio.ufpa.br/jspui/handle/2011/15726 |
dc.language.iso.fl_str_mv |
eng |
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eng |
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BMC Genomics |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by/4.0/ |
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openAccess |
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BioMed Central Ltd |
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Reino Unido |
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BioMed Central Ltd |
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