Genetic Engineering of Bacteria Lab., Division of Bacteriology, Indian Veterinary Research Institute, Izatnagar, India.
Mol Cell Probes. 2010 Jun;24(3):142-5. doi: 10.1016/j.mcp.2009.12.002. Epub 2009 Dec 16.
Brucellosis is a zoonosis of both public health and economic importance in many developing countries including India. Early detection and segregation of the infected animals are important in order to control the disease. Serodiagnostic tests for brucellosis is mainly based on detection of antibodies developed against lipopolysaccharide (LPS) component of cell. In this study we evaluated a protein antigen, 28 kDa outer membrane protein (OMP28), of Brucella melitensis as an alternative to LPS. Recombinant OMP28 was produced in Escherichia coli system. The efficacy of purified OMP28 was studied in an indirect enzyme-linked immunosorbent assay (ELISA) for diagnosis of brucellosis in field sera collected from different regions of country. Using known negative and known positive serum samples it was found that OMP28 is immunoreactive to Brucella infected cattle, sheep, goat and dog sera. Three hundred and eighty two cattle sera were screened by OMP28 antigen-based ELISA and the results were compared to rose Bengal plate agglutination Test (RBPT). Recombinant OMP28 antigen-based ELISA has shown sensitivity of 88.7%, specificity of 93.8% and accuracy of 92.9%. It was concluded that recombinant B. melitensis OMP28 could be used as a protein antigen for diagnosis of brucellosis in domestic animals.
布鲁氏菌病是一种人畜共患的疾病,在包括印度在内的许多发展中国家都具有公共卫生和经济重要性。早期发现和隔离受感染的动物对于控制疾病非常重要。布鲁氏菌病的血清诊断测试主要基于针对细胞脂多糖(LPS)成分产生的抗体的检测。在这项研究中,我们评估了一种蛋白质抗原,即 28 kDa 外膜蛋白(OMP28),作为 LPS 的替代品。重组 OMP28 在大肠杆菌系统中产生。在间接酶联免疫吸附试验(ELISA)中,研究了纯化的 OMP28 对来自该国不同地区的现场血清中布鲁氏菌病的诊断效果。使用已知的阴性和阳性血清样本发现,OMP28 与感染牛、羊、山羊和狗的血清具有免疫反应性。用 OMP28 抗原 ELISA 对 382 份牛血清进行了筛查,并将结果与孟加拉红平板凝集试验(RBPT)进行了比较。重组 OMP28 抗原 ELISA 的灵敏度为 88.7%,特异性为 93.8%,准确性为 92.9%。结论是,重组 B. melitensis OMP28 可作为诊断家畜布鲁氏菌病的蛋白质抗原。