Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams
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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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Elsevier
2021
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| Online Access: | 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 |
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| author | 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_browse | Carvalho Junior, Custodio A. Costa, Erica A. Gouveia, Aurora M.G. Lage, Andrey P. Moustacas, Valeria S. Paixao, Tatiane A. Robles, Carlos Alejandro Santos, Renato L. Sant’Anna, Felipe M. Silva, Teane M.A. Tsolis, Renee M. 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_sort | Xavier, Mariana N. |
| collection | INTA Digital |
| 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. |
| format | info:ar-repo/semantics/artículo |
| id | INTA10522 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA105222021-10-19T11:39:23Z Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams 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. Brucella Brucella Ovis Epididimitis Epididimitis Infecciosa Ovina PCR Epididymitis Ovine Infectious Epididymitis Rams Morueco Carneros Región Patagónica 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 2021-10-19T11:33:58Z 2021-10-19T11:33:58Z 2010 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Veterinary Microbiology 145 (1-2) : 158-164 (Septiembre 2010) |
| spellingShingle | Brucella Brucella Ovis Epididimitis Epididimitis Infecciosa Ovina PCR Epididymitis Ovine Infectious Epididymitis Rams Morueco Carneros Región Patagónica 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. 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 |
| 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_short | 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 |
| topic | Brucella Brucella Ovis Epididimitis Epididimitis Infecciosa Ovina PCR Epididymitis Ovine Infectious Epididymitis Rams Morueco Carneros Región Patagónica |
| 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 |
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