Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams

Autores
Xavier, Mariana N.; Silva, Teane M.A.; Costa, Erica A.; Paixao, Tatiane A.; Moustacas, Valeria S.; Carvalho Junior, Custodio A.; Sant’Anna, Felipe M.; Robles, Carlos Alejandro; Gouveia, Aurora M.G.; Lage, Andrey P.; Tsolis, Renee M.; Santos, Renato L.
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Brucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. The goal of this study was to develop a PCR test targeting specific B. ovis genomic sequences. Specific primer pairs were designed targeting 12 of those ORFs. Samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams (n = 9) every other week up to 180 days post infection (dpi), when tissue samples were obtained. Blood, serum, semen, urine, and preputial wash samples were obtained, in weekly intervals for 1 month, from eight rams belonging to a B. ovis-free flock. Semen samples were also obtained from rams belonging to naturally infected flocks (n = 40). The limit of detection of this PCR protocol was 100, 10, and 1 CFU/mL for semen, urine and prepucial wash samples, respectively. Sensitivity and specificity values obtained with this PCR method were similar to that of bacteriology when evaluating biological samples. Agreement between PCR and bacteriology results was greater than 90%. These results clearly indicate that this speciesspecific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.
Estación Experimental Agropecuaria Bariloche
Fil: Xavier, Mariana N. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Silva, Teane M.A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Costa, Erica A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Paixao, Tatiane A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Moustacas, Valeria S. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Carvalho Junior, Custodio A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Sant’Anna, Felipe M. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Grupo de Sanidad Animal; Argentina
Fil: Gouveia, Aurora M.G. Universidade Federal de Minas Gerais. Escola de Veterinária. Departamento de Medicina Veterinaria Preventiva; Brasil
Fil: Lage, Andrey P. Universidade Federal de Minas Gerais. Escola de Veterinária. Departamento de Medicina Veterinaria Preventiva; Brasil
Fil: Tsolis, Renee M. University of California. Department of Medical Microbiology and Immunology; Estados Unidos
Fil: Santos, Renato L. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fuente
Veterinary Microbiology 145 (1-2) : 158-164 (Septiembre 2010)
Materia
Brucella
Brucella Ovis
Epididimitis
Epididimitis Infecciosa Ovina
PCR
Epididymitis
Ovine Infectious Epididymitis
Rams
Morueco
Carneros
Región Patagónica
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/10522
Acceder
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oai_identifier_str oai:localhost:20.500.12123/10522
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
spelling Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in ramsXavier, Mariana N.Silva, Teane M.A.Costa, Erica A.Paixao, Tatiane A.Moustacas, Valeria S.Carvalho Junior, Custodio A.Sant’Anna, Felipe M.Robles, Carlos AlejandroGouveia, Aurora M.G.Lage, Andrey P.Tsolis, Renee M.Santos, Renato L.BrucellaBrucella OvisEpididimitisEpididimitis Infecciosa OvinaPCREpididymitisOvine Infectious EpididymitisRamsMoruecoCarnerosRegión PatagónicaBrucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. The goal of this study was to develop a PCR test targeting specific B. ovis genomic sequences. Specific primer pairs were designed targeting 12 of those ORFs. Samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams (n = 9) every other week up to 180 days post infection (dpi), when tissue samples were obtained. Blood, serum, semen, urine, and preputial wash samples were obtained, in weekly intervals for 1 month, from eight rams belonging to a B. ovis-free flock. Semen samples were also obtained from rams belonging to naturally infected flocks (n = 40). The limit of detection of this PCR protocol was 100, 10, and 1 CFU/mL for semen, urine and prepucial wash samples, respectively. Sensitivity and specificity values obtained with this PCR method were similar to that of bacteriology when evaluating biological samples. Agreement between PCR and bacteriology results was greater than 90%. These results clearly indicate that this speciesspecific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.Estación Experimental Agropecuaria BarilocheFil: Xavier, Mariana N. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Silva, Teane M.A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Costa, Erica A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Paixao, Tatiane A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Moustacas, Valeria S. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Carvalho Junior, Custodio A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Sant’Anna, Felipe M. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilFil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Grupo de Sanidad Animal; ArgentinaFil: Gouveia, Aurora M.G. Universidade Federal de Minas Gerais. Escola de Veterinária. Departamento de Medicina Veterinaria Preventiva; BrasilFil: Lage, Andrey P. Universidade Federal de Minas Gerais. Escola de Veterinária. Departamento de Medicina Veterinaria Preventiva; BrasilFil: Tsolis, Renee M. University of California. Department of Medical Microbiology and Immunology; Estados UnidosFil: Santos, Renato L. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; BrasilElsevier2021-10-19T11:33:58Z2021-10-19T11:33:58Z2010info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/10522https://www.sciencedirect.com/science/article/pii/S037811351000132X0378-1135https://doi.org/10.1016/j.vetmic.2010.02.037Veterinary Microbiology 145 (1-2) : 158-164 (Septiembre 2010)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccess2025-09-29T13:45:22Zoai:localhost:20.500.12123/10522instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:45:22.792INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
title Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
spellingShingle Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
Xavier, Mariana N.
Brucella
Brucella Ovis
Epididimitis
Epididimitis Infecciosa Ovina
PCR
Epididymitis
Ovine Infectious Epididymitis
Rams
Morueco
Carneros
Región Patagónica
title_short Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
title_full Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
title_fullStr Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
title_full_unstemmed Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
title_sort Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
dc.creator.none.fl_str_mv Xavier, Mariana N.
Silva, Teane M.A.
Costa, Erica A.
Paixao, Tatiane A.
Moustacas, Valeria S.
Carvalho Junior, Custodio A.
Sant’Anna, Felipe M.
Robles, Carlos Alejandro
Gouveia, Aurora M.G.
Lage, Andrey P.
Tsolis, Renee M.
Santos, Renato L.
author Xavier, Mariana N.
author_facet Xavier, Mariana N.
Silva, Teane M.A.
Costa, Erica A.
Paixao, Tatiane A.
Moustacas, Valeria S.
Carvalho Junior, Custodio A.
Sant’Anna, Felipe M.
Robles, Carlos Alejandro
Gouveia, Aurora M.G.
Lage, Andrey P.
Tsolis, Renee M.
Santos, Renato L.
author_role author
author2 Silva, Teane M.A.
Costa, Erica A.
Paixao, Tatiane A.
Moustacas, Valeria S.
Carvalho Junior, Custodio A.
Sant’Anna, Felipe M.
Robles, Carlos Alejandro
Gouveia, Aurora M.G.
Lage, Andrey P.
Tsolis, Renee M.
Santos, Renato L.
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Brucella
Brucella Ovis
Epididimitis
Epididimitis Infecciosa Ovina
PCR
Epididymitis
Ovine Infectious Epididymitis
Rams
Morueco
Carneros
Región Patagónica
topic Brucella
Brucella Ovis
Epididimitis
Epididimitis Infecciosa Ovina
PCR
Epididymitis
Ovine Infectious Epididymitis
Rams
Morueco
Carneros
Región Patagónica
dc.description.none.fl_txt_mv Brucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. The goal of this study was to develop a PCR test targeting specific B. ovis genomic sequences. Specific primer pairs were designed targeting 12 of those ORFs. Samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams (n = 9) every other week up to 180 days post infection (dpi), when tissue samples were obtained. Blood, serum, semen, urine, and preputial wash samples were obtained, in weekly intervals for 1 month, from eight rams belonging to a B. ovis-free flock. Semen samples were also obtained from rams belonging to naturally infected flocks (n = 40). The limit of detection of this PCR protocol was 100, 10, and 1 CFU/mL for semen, urine and prepucial wash samples, respectively. Sensitivity and specificity values obtained with this PCR method were similar to that of bacteriology when evaluating biological samples. Agreement between PCR and bacteriology results was greater than 90%. These results clearly indicate that this speciesspecific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.
Estación Experimental Agropecuaria Bariloche
Fil: Xavier, Mariana N. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Silva, Teane M.A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Costa, Erica A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Paixao, Tatiane A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Moustacas, Valeria S. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Carvalho Junior, Custodio A. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Sant’Anna, Felipe M. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
Fil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Grupo de Sanidad Animal; Argentina
Fil: Gouveia, Aurora M.G. Universidade Federal de Minas Gerais. Escola de Veterinária. Departamento de Medicina Veterinaria Preventiva; Brasil
Fil: Lage, Andrey P. Universidade Federal de Minas Gerais. Escola de Veterinária. Departamento de Medicina Veterinaria Preventiva; Brasil
Fil: Tsolis, Renee M. University of California. Department of Medical Microbiology and Immunology; Estados Unidos
Fil: Santos, Renato L. Universidade Federal de Minas. Escola de Veterinária. Departamento de Clínica e Cirurgia Veterinária; Brasil
description Brucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. The goal of this study was to develop a PCR test targeting specific B. ovis genomic sequences. Specific primer pairs were designed targeting 12 of those ORFs. Samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams (n = 9) every other week up to 180 days post infection (dpi), when tissue samples were obtained. Blood, serum, semen, urine, and preputial wash samples were obtained, in weekly intervals for 1 month, from eight rams belonging to a B. ovis-free flock. Semen samples were also obtained from rams belonging to naturally infected flocks (n = 40). The limit of detection of this PCR protocol was 100, 10, and 1 CFU/mL for semen, urine and prepucial wash samples, respectively. Sensitivity and specificity values obtained with this PCR method were similar to that of bacteriology when evaluating biological samples. Agreement between PCR and bacteriology results was greater than 90%. These results clearly indicate that this speciesspecific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.
publishDate 2010
dc.date.none.fl_str_mv 2010
2021-10-19T11:33:58Z
2021-10-19T11:33:58Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/10522
https://www.sciencedirect.com/science/article/pii/S037811351000132X
0378-1135
https://doi.org/10.1016/j.vetmic.2010.02.037
url http://hdl.handle.net/20.500.12123/10522
https://www.sciencedirect.com/science/article/pii/S037811351000132X
https://doi.org/10.1016/j.vetmic.2010.02.037
identifier_str_mv 0378-1135
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Veterinary Microbiology 145 (1-2) : 158-164 (Septiembre 2010)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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score 12.559606

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