Canine babesiosis, a tick-borne haemoprotozoan disease of dogs, is of significance globally due to its rapid spread. A precise confirmatory diagnosis is required to curtail the rapid spread of infection. Our study described the evaluation of recombinant BgSA3 protein based indirect ELISA for sero-diagnosis and sero-surveillance of Babesia gibsoni infection in dogs. A partial BgSA3 gene segment (1921 bp) of B. gibsoni, encoding for recombinant truncated BgSA3 (75 kDa) protein devoid of predicted signal peptide (23 aa) at N-terminus and transmembrane region (20 aa) at C-terminus, was expressed in E. coli using a pET28a(+) vector. The rBgSA3 protein purified under native conditions using Ni-NTA superflow cartridge was confirmed by SDS-PAGE and Western blotting using sera from dogs infected/uninfected with B. gibsoni, and erythrocyte lysate/ plasma from infected/uninfected dogs. The rBgSA3 protein was specific only to B. gibsoni antibodies but did not react with uninfected sera. Further, rBgSA3 protein was evaluated for sero-diagnosis/sero-surveillance using Indirect-ELISA format. There was no cross reactivity to B. vogeli, E. canis, H. canis and D. repens infected dogs serum samples. The diagnostic sensitivity and specificity of rBgSA3 based I-ELISA was found to be 86.4 and 93.1 % respectively, in comparison with cytb based PCR assay. Additionally, rBgSA3-ELISA evaluated using survey serum samples (n = 287), detected 11.85 % samples as positive. In conclusion, B. gibsoni infection, an emerging disease is prevalent in the present study area and the standardized rBgSA3 protein based indirect-ELISA was found to be a specific and sensitive test for large scale sero-diagnosis and sero-surveillance of B. gibsoni infection in dogs.

犬幼犬病是由tick传播的犬原虫病，由于其迅速传播而在全球具有重要意义。需要精确的确认性诊断来减少感染的迅速传播。我们的研究描述了基于重组BgSA3蛋白的间接ELISA对犬巴贝斯吉布森犬的血清学诊断和血清学监测的评估。的部分BgSA3基因区段（1921 bp）的B. gibsoni，编码重组截短BgSA3（75 kDa）的在在C末端N末端和跨膜区（20个氨基酸）预测的信号肽（23个氨基酸）的蛋白缺乏，是在大肠杆菌中表达使用pET28a（+）向量。在自然条件下使用Ni-NTA超流小柱纯化的rBgSA3蛋白通过SDS-PAGE和Western blotting证实，使用的血清来自来自感染或未感染长臂猿的狗的血清以及来自感染/未感染的狗的红细胞裂解液/血浆。rBgSA3蛋白仅特异于长双歧杆菌抗体，但不与未感染的血清反应。此外，使用间接ELISA形式评估rBgSA3蛋白的血清诊断/血清监测。与B. vogeli，E. canis，H. canis和D. repens没有交叉反应。被感染的狗的血清样本。与基于cytb的PCR分析相比，基于rBgSA3的I-ELISA的诊断敏感性和特异性分别为86.4％和93.1％。另外，使用调查血清样本（n = 287）评估了rBgSA3-ELISA，检测到11.85％的样本为阳性。综上所述，吉本氏杆菌感染是一种正在出现的疾病，是当前研究领域中普遍存在的问题，基于标准化的基于rBgSA3蛋白的间接ELISA被认为是针对吉本氏杆菌的大规模血清学诊断和血清学监测的特异性和灵敏性测试。狗感染。