Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans

ABSTRACT: laboratory diagnosis of canine brucellosis includes serological and bacteriological tests; the blood culture is considered the gold standard, but it presents issues of sensitivity and delay in results. Therefore, the polymerase chain reaction (PCR) could be useful to detect low amounts of...

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Autores:
Castrillón Salazar, Laura Lucía
Sánchez Jiménez, Miryan Margot
Ortiz Román, Luisa Fernanda
Giraldo Echeverri, Carlos Andrés
Olivera Ángel, Martha
Tipo de recurso:
Article of investigation
Fecha de publicación:
2014
Institución:
Universidad de Antioquia
Repositorio:
Repositorio UdeA
Idioma:
eng
OAI Identifier:
oai:bibliotecadigital.udea.edu.co:10495/8315
Acceso en línea:
http://hdl.handle.net/10495/8315
Palabra clave:
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openAccess
License
https://creativecommons.org/licenses/by-nc-sa/2.5/co/
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dc.title.spa.fl_str_mv Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
dc.title.translated.spa.fl_str_mv Aplicación de una prueba de reacción en cadena de la polimerasa para detección de Brucella canis a partir de muestras clínicas de caninos y humanos
title Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
spellingShingle Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
title_short Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
title_full Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
title_fullStr Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
title_full_unstemmed Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
title_sort Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans
dc.creator.fl_str_mv Castrillón Salazar, Laura Lucía
Sánchez Jiménez, Miryan Margot
Ortiz Román, Luisa Fernanda
Giraldo Echeverri, Carlos Andrés
Olivera Ángel, Martha
dc.contributor.author.none.fl_str_mv Castrillón Salazar, Laura Lucía
Sánchez Jiménez, Miryan Margot
Ortiz Román, Luisa Fernanda
Giraldo Echeverri, Carlos Andrés
Olivera Ángel, Martha
dc.contributor.researchgroup.spa.fl_str_mv Biogénesis
Grupo Vericel
description ABSTRACT: laboratory diagnosis of canine brucellosis includes serological and bacteriological tests; the blood culture is considered the gold standard, but it presents issues of sensitivity and delay in results. Therefore, the polymerase chain reaction (PCR) could be useful to detect low amounts of bacterial DNA from clinical samples and provide results within hours. Objective: to evaluate the sensitivity and specificity of PCR for the detection of Brucella canis in whole blood samples. Methods: blood samples from 499 dogs from kennels in two Colombian regions and 91 co-inhabiting humans were used. The 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) from serum and blood culture and PCR tests from whole blood were performed on all samples. Bayes theorem was used to establish the sensitivity and specificity of the PCR test compared with the other tests performed. Results: 9.9% of the evaluated co-inhabiting humans yielded positive serological results and 0% were positive by PCR or blood culture tests. 10.8% of dog samples were positive by blood culture, 19% were positive by PCR and 13% were positive by 2ME-RSAT. 7% of the samples were positive by all tests. Compared with blood culture, PCR had a sensitivity of 92.6% and a specificity of 90% for canine samples. Compared with 2ME-RSAT, it had a sensitivity of 77.4% and a specificity of 89.2%. When PCR and 2ME-RSAT results were compared with blood culture, a higher number of positive samples were retrieved than when results of only individual tests were applied. Conclusions: PCR is useful to detect B. canis in clinical samples; however, it is preferable to include the 2ME-RSAT test, as this improves the accuracy of the diagnosis. The PCR results are obtained within 24 to 48 hours and do not require the presence of whole bacterial cells to detect DNA.
publishDate 2014
dc.date.issued.none.fl_str_mv 2014
dc.date.accessioned.none.fl_str_mv 2017-09-25T20:57:51Z
dc.date.available.none.fl_str_mv 2017-09-25T20:57:51Z
dc.type.spa.fl_str_mv Artículo de investigación
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dc.identifier.citation.spa.fl_str_mv Sánchez-Jiménez MM, Ortiz-Román LF, Castrillón-Salazar LL, Giraldo-Echeverri CA, Olivera-Angel M. Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans. Rev. Colomb. Cienc. Pecu. 2014; 27(1): 3-11
dc.identifier.issn.none.fl_str_mv 0120-0690
dc.identifier.uri.none.fl_str_mv http://hdl.handle.net/10495/8315
dc.identifier.eissn.none.fl_str_mv 2256-2958
identifier_str_mv Sánchez-Jiménez MM, Ortiz-Román LF, Castrillón-Salazar LL, Giraldo-Echeverri CA, Olivera-Angel M. Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans. Rev. Colomb. Cienc. Pecu. 2014; 27(1): 3-11
0120-0690
2256-2958
url http://hdl.handle.net/10495/8315
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.ispartofjournalabbrev.spa.fl_str_mv Rev. Colomb. Cienc. Pecu.
dc.relation.citationendpage.spa.fl_str_mv 11
dc.relation.citationissue.spa.fl_str_mv 1
dc.relation.citationstartpage.spa.fl_str_mv 3-11
dc.relation.citationvolume.spa.fl_str_mv 27
dc.relation.ispartofjournal.spa.fl_str_mv Revista Colombiana de Ciencias Pecuarias
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dc.publisher.spa.fl_str_mv Universidad de Antioquia, Facultad de Ciencias Agrarias
dc.publisher.place.spa.fl_str_mv Medellín, Colombia
institution Universidad de Antioquia
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spelling Castrillón Salazar, Laura LucíaSánchez Jiménez, Miryan MargotOrtiz Román, Luisa FernandaGiraldo Echeverri, Carlos AndrésOlivera Ángel, MarthaBiogénesisGrupo Vericel2017-09-25T20:57:51Z2017-09-25T20:57:51Z2014Sánchez-Jiménez MM, Ortiz-Román LF, Castrillón-Salazar LL, Giraldo-Echeverri CA, Olivera-Angel M. Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humans. Rev. Colomb. Cienc. Pecu. 2014; 27(1): 3-110120-0690http://hdl.handle.net/10495/83152256-2958ABSTRACT: laboratory diagnosis of canine brucellosis includes serological and bacteriological tests; the blood culture is considered the gold standard, but it presents issues of sensitivity and delay in results. Therefore, the polymerase chain reaction (PCR) could be useful to detect low amounts of bacterial DNA from clinical samples and provide results within hours. Objective: to evaluate the sensitivity and specificity of PCR for the detection of Brucella canis in whole blood samples. Methods: blood samples from 499 dogs from kennels in two Colombian regions and 91 co-inhabiting humans were used. The 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) from serum and blood culture and PCR tests from whole blood were performed on all samples. Bayes theorem was used to establish the sensitivity and specificity of the PCR test compared with the other tests performed. Results: 9.9% of the evaluated co-inhabiting humans yielded positive serological results and 0% were positive by PCR or blood culture tests. 10.8% of dog samples were positive by blood culture, 19% were positive by PCR and 13% were positive by 2ME-RSAT. 7% of the samples were positive by all tests. Compared with blood culture, PCR had a sensitivity of 92.6% and a specificity of 90% for canine samples. Compared with 2ME-RSAT, it had a sensitivity of 77.4% and a specificity of 89.2%. When PCR and 2ME-RSAT results were compared with blood culture, a higher number of positive samples were retrieved than when results of only individual tests were applied. Conclusions: PCR is useful to detect B. canis in clinical samples; however, it is preferable to include the 2ME-RSAT test, as this improves the accuracy of the diagnosis. The PCR results are obtained within 24 to 48 hours and do not require the presence of whole bacterial cells to detect DNA.RESUMEN: el diagnóstico de laboratorio de brucelosis canina incluye pruebas serológicas y bacteriológicas. El hemocultivo se considera el estándar de oro, pero tiene problemas de sensibilidad y tiempo de entrega de los resultados. Por eso, la reacción en cadena de la polimerasa (PCR) podría ser útil para detectar pequeñas cantidades de ADN bacteriano de muestras clínicas y proporcionar resultados en horas. Objetivo: evaluar la sensibilidad y especificidad de una PCR para la detección de Brucella canis, en muestras de sangre total de 499 perros de perreras y 91 humanos cohabitantes de dos regiones de Colombia. Métodos: se realizaron pruebas de aglutinación rápida en placa con 2-mercaptoetanol (2ME-PARP) a partir de suero, PCR y hemocultivo a partir de sangre total. El teorema de Bayes se utilizó para establecer la sensibilidad y especificidad de la prueba de PCR respecto a las demás. Resultados: 9,9% de los humanos cohabitantes evaluados resultaron positivos para la prueba serológica, y el 0% fue positivo para PCR o hemocultivo. 10,8% de las muestras de perros fueron positivas para hemocultivo, 19% fueron positivas para PCR y 13% eran 2ME-PARP positivo. 7% de las muestras fueron positivas para todos los ensayos. En las muestras caninas, la PCR tuvo una sensibilidad del 92,6% y una especificidad del 90% en comparación con el hemocultivo. En comparación con 2ME-PARP, tuvo una sensibilidad del 77,4% y una especificidad del 89,2%. Cuando se compararon los resultados de la PCR y 2ME-PARP con el hemocultivo, un mayor número de muestras positivas fueron obtenidas que usando los resultados de cada una. Conclusiones: la PCR es útil para detectar B.canis en muestras clínicas, sin embargo, es recomendable incluir la prueba de 2ME-PARP, lo que mejora la exactitud del diagnóstico, se obtienen los resultados en 24 ó 48 horas y no requieren la presencia de células bacterianas completas para detectar ADN.COL0066561COL0078089application/pdfengUniversidad de Antioquia, Facultad de Ciencias AgrariasMedellín, Colombiahttps://creativecommons.org/licenses/by-nc-sa/2.5/co/https://creativecommons.org/licenses/by-nc-sa/4.0/Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Application of a polymerase chain reaction test for the detection of Brucella canis from clinical samples of canines and humansAplicación de una prueba de reacción en cadena de la polimerasa para detección de Brucella canis a partir de muestras clínicas de caninos y humanosArtículo de investigaciónhttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARThttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionRev. Colomb. Cienc. 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