Instituto de Patobiologia, IP-IPVet, UEDD INTA-Conicet, N Repetto y de Los Reseros s/n, Hurlingham B1686, Buenos Aires, Argentina; IP-IPVet, UEDD INTA-Conicet, N Repetto y de Los Reseros s/n, Hurlingham B1686, Buenos Aires, Argentina.
IP-IPVet, UEDD INTA-Conicet, N Repetto y de Los Reseros s/n, Hurlingham B1686, Buenos Aires, Argentina.
Res Vet Sci. 2022 Oct;147:1-6. doi: 10.1016/j.rvsc.2022.03.015. Epub 2022 Mar 17.
Different fractions of Brucella (B) abortus or Brucella melitensis have been used as antigens for the detection of anti-Brucella antibodies in goat sera, being their accomplishment cumbersome and time consuming. In an attempt to achieve a simpler enzyme-linked immunosorbent assay (ELISA) antigen preparation method for serodiagnosis of caprine brucellosis, we developed and evaluated a B. melitensis whole-cell lysate antigen-based indirect ELISA (Bm-WCL iELISA). A total of 162 serum samples from female crossbred goats collected from non-vaccinated herds against brucellosis were classified according to the buffered plate antigen (BPA) screening test and the complement fixation (CF) test and used for the indirect ELISA (iELISA) evaluation. The Bm-WCL iELISA showed a high Se and Sp [95.7% (CI 88.1% - 98.8%), and 92.4% (CI 83.4% - 96.7%), respectively] to detect the serological response against Brucella in commercial goat herds, and an almost perfect agreement with combined official tests results (κ = 0.88), when goat sera with concordant results in both official serological tests (BPA and CF; n = 136) were used. However, the agreement dropped to substantial (k > 0.73) when 26 goat serum samples with BPA and CF not concordant results were incorporated for the iELISA performance evaluation and the comparison was made for each test independently. Comparison of the Bm-WCL iELISA results with Brucella abortus sLPS iELISA showed almost perfect agreement (κ > 0.83). Even when a larger number of samples are needed to validate this test, these preliminary results encourage the optimization of the Brucella melitensis whole cell lysate antigen-based iELISA.
不同的布鲁氏菌（B） abortus 或布鲁氏菌 melitensis 片段已被用作检测山羊血清中抗布鲁氏菌抗体的抗原，其实现过程繁琐且耗时。为了实现一种更简单的酶联免疫吸附试验（ELISA）抗原制备方法，用于诊断山羊布鲁氏菌病，我们开发并评估了一种基于布鲁氏菌 melitensis 全细胞裂解物抗原的间接 ELISA（Bm-WCL iELISA）。共收集了来自未接种布鲁氏菌病疫苗的杂交母羊的 162 份血清样本，根据缓冲平板抗原（BPA）筛选试验和补体结合（CF）试验进行分类，并用于间接 ELISA（iELISA）评估。Bm-WCL iELISA 对商业山羊群中针对布鲁氏菌的血清学反应显示出高的 Se 和 Sp [95.7%（置信区间 88.1% - 98.8%）和 92.4%（置信区间 83.4% - 96.7%）]，与联合官方检测结果几乎完全一致（κ=0.88），当使用两种官方血清学检测（BPA 和 CF；n=136）结果一致的山羊血清时。然而，当纳入 26 份 BPA 和 CF 结果不一致的山羊血清样本用于 iELISA 性能评估时，一致性降至高度（k>0.73），并且对每个测试分别进行了比较。Bm-WCL iELISA 结果与布鲁氏菌 abortus sLPS iELISA 的比较显示出几乎完全一致（κ>0.83）。即使需要更多的样本来验证该测试，这些初步结果也鼓励优化基于布鲁氏菌 melitensis 全细胞裂解物抗原的 iELISA。
