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Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams.
Vet Microbiol. 2010 Sep 28; 145(1-2):158-64.VM

Abstract

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 species-specific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.

Authors+Show Affiliations

Departamento de Clínica e Cirurgia Veterinária, Escola de Veterinária, Universidade Federal de Minas Gerais, 31270-901 Belo Horizonte, MG, Brazil.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20347534

Citation

Xavier, Mariana N., et al. "Development and Evaluation of a Species-specific PCR Assay for the Detection of Brucella Ovis Infection in Rams." Veterinary Microbiology, vol. 145, no. 1-2, 2010, pp. 158-64.
Xavier MN, Silva TM, Costa EA, et al. Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams. Vet Microbiol. 2010;145(1-2):158-64.
Xavier, M. N., Silva, T. M., Costa, E. A., Paixão, T. A., Moustacas, V. S., Carvalho, C. A., Sant'Anna, F. M., Robles, C. A., Gouveia, A. M., Lage, A. P., Tsolis, R. M., & Santos, R. L. (2010). Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams. Veterinary Microbiology, 145(1-2), 158-64. https://doi.org/10.1016/j.vetmic.2010.02.037
Xavier MN, et al. Development and Evaluation of a Species-specific PCR Assay for the Detection of Brucella Ovis Infection in Rams. Vet Microbiol. 2010 Sep 28;145(1-2):158-64. PubMed PMID: 20347534.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and evaluation of a species-specific PCR assay for the detection of Brucella ovis infection in rams. AU - Xavier,Mariana N, AU - Silva,Teane M A, AU - Costa,Erica A, AU - Paixão,Tatiane A, AU - Moustacas,Valéria S, AU - Carvalho,Custódio A,Jr AU - Sant'Anna,Felipe M, AU - Robles,Carlos A, AU - Gouveia,Aurora M G, AU - Lage,Andrey P, AU - Tsolis,Renée M, AU - Santos,Renato L, Y1 - 2010/03/07/ PY - 2009/12/16/received PY - 2010/02/26/revised PY - 2010/02/26/accepted PY - 2010/3/30/entrez PY - 2010/3/30/pubmed PY - 2010/12/18/medline SP - 158 EP - 64 JF - Veterinary microbiology JO - Vet. Microbiol. VL - 145 IS - 1-2 N2 - 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 species-specific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams. SN - 1873-2542 UR - https://www.unboundmedicine.com/medline/citation/20347534/Development_and_evaluation_of_a_species_specific_PCR_assay_for_the_detection_of_Brucella_ovis_infection_in_rams_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-1135(10)00132-X DB - PRIME DP - Unbound Medicine ER -