Comparison of 2 ELISAs for detecting exposure to Brucella ovis.

Control of Brucella ovis infection in sheep flocks in the United States depends on early detection of B. ovis antibodies via serologic testing. We used 2,276 sheep sera and various cutoff values to compare seroprevalence and agreement between 2 ELISAs: the National Veterinary Services Laboratories (NVSL) B. ovis indirect ELISA and the IDEXX B. ovis ELISA kit. A subset of 295 sera was used to compare agreement and evaluate relative sensitivity and specificity of the 2 ELISAs with an agar gel immunodiffusion (AGID) test kit. There was no significant difference in B. ovis seroprevalence between the ELISAs; however, there was poor agreement between them. When the AGID test was used as the reference test, the IDEXX ELISA with a moderate cutoff value (S/P ratio = 45%) had the highest relative sensitivity of 38.1% and specificity of 92.0%. The NVSL ELISA with a lax cutoff value (S/P ratio = 0.75) had relative sensitivity of 19.1% and specificity of 94.6%. Receiver operating characteristic analysis revealed that optimal cutoff values for the NVSL and IDEXX ELISAs were 0.091 and 16.5%, respectively. This results in sensitivity and specificity of 85.7% and 31.8% for the NVSL ELISA, and sensitivity and specificity of 81.0% and 53.6% for the IDEXX ELISA, respectively.

Seroprevalence and risk factors of Brucella ovis in domestic sheep in Wyoming, USA.

BACKGROUND: Brucella ovis causes a sexually transmitted, infectious disease of domestic sheep characterized by genital lesions and epididymitis in rams, placentitis and rare abortions in ewes, and neonatal mortality in lambs. This study was designed to 1) estimate animal and flock seroprevalence of B. ovis in sheep across Wyoming, USA, and 2) describe epidemiologic risk factors associated with seropositive sheep and flocks. For the animal seroprevalence estimate, 2423 blood samples were collected from sheep on 18 producer-selected operations and a questionnaire about possible risk factors was distributed. For the flock seroprevalence estimate, blood samples from 82 operations were obtained, including samples from the previous 18 operations and 64 additional operations that sent samples to the Wyoming State Veterinary Laboratory for diagnostic testing. Categorical risk factors were created based on questionnaires and submission forms. Sera was analyzed using the B. ovis enzyme-linked immunosorbent assay. RESULTS: Estimated true animal and flock seroprevalence were 0.53% (95% CI: 0.21-1.01%; 22/2,423) and 22.5% (95% CI: 14-32%; 18/82), respectively. Using Fisher's exact and Mid-p exact tests to compare apparent seroprevalence with respect to possible risk factors, increased age and breed type were risk factors associated with seropositive sheep, while region and large flock size were risk factors associated with seropositive flocks. CONCLUSIONS: Results from this study suggest few sheep have been exposed to B. ovis, but many flocks contain at least one seropositive animal. Each region in Wyoming contained at least one seropositive animal and flock, emphasizing the importance of disease-free documentation before purchasing new sheep. Aged sheep (≥ 6 years of age) had the highest seroprevalence among age groups; hence, we propose the separation of young rams from older rams to help reduce disease spread outside the breeding season. Wool breeds (Rambouillet and Merino) may be less susceptible to B. ovis infection given they had the lowest animal seroprevalence of the breed types, and large flocks (> 100 breeding rams) had the highest seroprevalence of the flock size categories, likely due to more intensive management strategies that can contribute to the introduction and persistence of B. ovis infection in sheep and flocks.