Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University-Medical College, 31-008 Kraków, Poland.
Bioidea Company, 02-991 Warsaw, Poland.
Int J Mol Sci. 2023 Mar 31;24(7):6569. doi: 10.3390/ijms24076569.
Bovine mastitis is the most common disease affecting dairy cattle worldwide and it generates substantial losses for cattle breeders. One of the most common pathogens identified in infected milk samples is . Currently, there is no fast test for recognizing bacteria species on the market. The aim of this study was to bioinformatically and laboratory detect and characterize the fibronectin binding protein A (FnBPA) of (SA) in milk samples obtained from cows diagnosed with mastitis. More than 90,000,000 amino acid sequences were subjected to bioinformatic detection in the search for a potential biomarker for bovine SA. The analysis of FnBPA included the detection of signal peptides and nonclassical proteins, antigenicity, and the prediction of epitopes. To confirm the presence of the A gene in four SA isolates, amplification with specific primers was performed. FnBPA was detected by immunoblotting. The immunoreactivity and selectivity were performed with monoclonal anti-FnBPA antibodies and SA-negative serum. The bioinformatic analysis showed that FnBPA is a surface, conservative, immunoreactive, and species-specific protein with antigenic potential. Its presence was confirmed in all of the SA isolates we studied. Immunoblotting proved its immunoreactivity and specificity. Thus, it can be considered a potential biomarker in mastitis immunodiagnostics.
奶牛乳腺炎是全球范围内影响奶牛的最常见疾病，给奶牛养殖者造成了巨大的损失。在感染的奶样中最常见的病原体之一是 。目前，市场上还没有快速识别细菌种类的检测方法。本研究旨在通过生物信息学和实验室检测和鉴定从乳腺炎奶牛获得的奶样中的纤维连接蛋白结合蛋白 A（FnBPA）。在寻找牛 SA 的潜在生物标志物的研究中，对超过 90000000 个氨基酸序列进行了生物信息学检测。FnBPA 的分析包括信号肽和非经典蛋白、抗原性和表位预测的检测。为了确认四个 SA 分离株中 A 基因的存在，用特异性引物进行了扩增。通过免疫印迹检测 FnBPA。用单克隆抗 FnBPA 抗体和 SA 阴性血清进行了免疫反应性和选择性检测。生物信息学分析表明，FnBPA 是一种表面保守、免疫反应性和种特异性的蛋白，具有抗原潜力。在我们研究的所有 SA 分离株中都证实了其存在。免疫印迹证明了其免疫反应性和特异性。因此，它可以被认为是乳腺炎免疫诊断中的一个潜在的生物标志物。
