Brucellosis is a zoonotic disease transmitted to humans from infected animals. As a systemic disease, it can harm any organ or system of the host body. Human brucellosis presents with various clinical symptoms, which makes diagnosis challenging. Serological diagnosis of brucellosis is based on ELISA or agglutination tests, which use colorimetry to detect antibodies generated against lipopolysaccharide (LPS) or extracts from whole-cell bacteria. To construct a protein that can specifically recognize Brucella, we analyzed hydrophilicity, accessibility, flexibility, antigenicity, and β-turns using a protein network server. Then, we chose the most abundant immunodominant epitopes of the outer membrane proteins omp31, BP26, omp2b and omp16. Based on the sequences of these major epitopes, fifteen major immunodominant epitopes were selected to construct a synthetic Brucella recombinant multiepitope outer membrane protein (rOmp) gene. This recombinant gene was expressed in E. coli, and the produced protein was purified by Ni-NTA affinity purification. The purified protein was tested in an indirect ELISA assay, demonstrating a high level of sensitivity and specificity. This technique is creating a unique antigen that, coupled with overexpression and low-cost purification, offers a promising diagnosis of both human and animal brucellosis, with the potential to avoid the disadvantages of whole brucellosis-antigen-based assays.

布鲁氏菌病是一种从被感染的动物传播给人类的人畜共患疾病。作为一种全身性疾病，它可以损害宿主身体的任何器官或系统。人类布鲁氏菌病具有各种临床症状，这使诊断具有挑战性。布鲁氏菌病的血清学诊断基于ELISA或凝集试验，该试验使用比色法检测针对脂多糖（LPS）生成的抗体或全细胞细菌提取物。为了构建可以特异性识别布鲁氏菌的蛋白质，我们分析了亲水性，可及性，柔韧性，抗原性和β-使用蛋白质网络服务器打开。然后，我们选择了外膜蛋白omp31，BP26，omp2b和omp16的最丰富的免疫优势表位。基于这些主要表位的序列，选择了十五种主要的免疫显性表位来构建合成的布鲁氏菌重组多表位外膜蛋白（rOmp）基因。该重组基因在大肠杆菌中表达，然后通过Ni-NTA亲和纯化纯化产生的蛋白质。纯化的蛋白质在间接ELISA分析中进行了测试，证明了高水平的敏感性和特异性。这项技术正在创造一种独特的抗原，再加上过表达和低成本纯化，可为人类和动物布鲁氏菌病提供有希望的诊断，并有可能避免基于布鲁氏菌病抗原的整个检测方法的弊端。