Department of Parasitology, Faculty of Veterinary Medicine, University of Selcuk, Konya, Türkiye.
Jiangsu Key Laboratory for High-tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Agri-animal Husbandry Vocational College, Taizhou, China.
Front Cell Infect Microbiol. 2023 Aug 16;13:1238369. doi: 10.3389/fcimb.2023.1238369. eCollection 2023.
Ovine babesiosis caused by is an economically significant disease. Recently, a few -specific proteins, including recombinant secreted antigen-1 (rBoSA1), have been identified. Immunological analyses revealed that rBoSA1 resides within the cytoplasm of infected erythrocytes and exhibits robust antigenic properties for detecting anti- antibodies. This protein is released into the bloodstream during the parasite's development. It would be possible to diagnose active infections by detecting this secretory protein. For this purpose, a rBoSA1-specific polyclonal antibody-based sandwich ELISA was optimized in this study. Blood samples taken from the naturally (n: 100) and experimentally (n: 15) infected sheep were analyzed for the presence of native BoSA1. The results showed that native BoSA1 was detectable in 98% of naturally infected animals. There was a positive correlation between parasitemia level in microscopy and protein density in sandwich ELISA. Experimentally infected animals showed positive reactions from the first or second day of inoculations. However, experimental infections carried out by ticks revealed the native BoSA1 was detectable from the 7 day of tick attachment when the parasite began to be seen microscopically. Sandwich ELISA was sensitive enough to detect rBoSA1 protein at a 1.52 ng/ml concentration. Additionally, no serological cross-reactivity was observed between animals infected with various piroplasm species, including , , , and Taken collectively, the findings show that the rBoSA1-specific polyclonal antibody-based sandwich ELISA can be successfully used to diagnose clinical infections in sheep at the early stage.
绵羊巴贝斯虫病由 引起，是一种具有重要经济意义的疾病。最近，已经鉴定出一些 - 特异性蛋白，包括重组 分泌抗原-1（rBoSA1）。免疫分析表明，rBoSA1存在于感染的红细胞细胞质中，具有检测抗- 抗体的强抗原特性。这种蛋白质在寄生虫发育过程中释放到血液中。通过检测这种分泌蛋白，有可能诊断出活动性感染。为此，本研究优化了一种基于 rBoSA1 特异性多克隆抗体的夹心 ELISA。分析了来自自然（n：100）和实验（n：15）感染绵羊的血液样本中是否存在天然 BoSA1。结果表明，98%的自然感染动物可检测到天然 BoSA1。显微镜检查中的寄生虫血症水平与夹心 ELISA 中的蛋白密度之间存在正相关。从接种的第 1 天或第 2 天开始，实验感染动物就出现了阳性反应。然而，通过 蜱进行的实验感染表明，当寄生虫在显微镜下开始可见时，从蜱附着的第 7 天开始可以检测到天然 BoSA1。夹心 ELISA 足够灵敏，可检测到浓度为 1.52ng/ml 的 rBoSA1 蛋白。此外，在感染各种梨形虫的动物之间没有观察到血清学交叉反应，包括 、 、 和 。总的来说，这些发现表明，基于 rBoSA1 特异性多克隆抗体的夹心 ELISA 可成功用于诊断绵羊临床 感染的早期阶段。
