Rugdech P, Anuntagool N, Sirisinha S
Faculty of Science, Mahidol University, Chulabhorn Research Institute, Bangkok, Thailand.
Am J Trop Med Hyg. 1995 Mar;52(3):231-5. doi: 10.4269/ajtmh.1995.52.231.
Monoclonal antibodies (MAbs) specific for Pseudomonas pseudomallei antigens were produced by immunizing BALB/c mice with a crude whole cell extract. Hybrids secreting MAbs specific for P. pseudomallei antigens were identified by an indirect enzyme-linked immunosorbent assay (ELISA) against a panel of crude whole cell extracts from P. pseudomallei, P. cepacia, P. aeruginosa, P. putida, P. alcaligenes, Xanthomonas maltophilia, Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Salmonella typhi, S. krefeld, S. enteritidis, Proteus mirabilis, and Staphylococcus aureus. Of the six specific clones, clone 5F8, which was IgM-producing and which reacted with all 56 P. pseudomallei isolates, was selected for further characterization and evaluation of its possible diagnostic potential. Results obtained from the indirect ELISA against various P. pseudomallei antigens, from direct bacterial agglutination, and from immunofluorescence tests suggested that 5F8 reacted with the surface envelope, and probably specifically with an epitope of the lipopolysaccharide. The antibody could be readily used to identify P. pseudomallei in primary culture or in a simulated hemoculture. The antibody was also used to prepare an affinity-purified antigen for use in an indirect ELISA that was highly sensitive and specific for the detection of circulating antibody in patients with acute septicemic melioidosis.
用粗制全细胞提取物免疫BALB/c小鼠，制备针对类鼻疽杆菌抗原的单克隆抗体（MAbs）。通过间接酶联免疫吸附测定（ELISA），针对一组来自类鼻疽杆菌、洋葱伯克霍尔德菌、铜绿假单胞菌、恶臭假单胞菌、产碱假单胞菌、嗜麦芽窄食单胞菌、肺炎克雷伯菌、大肠埃希菌、阴沟肠杆菌、伤寒沙门菌、克雷费尔德沙门菌、肠炎沙门菌、奇异变形杆菌和金黄色葡萄球菌的粗制全细胞提取物，鉴定分泌针对类鼻疽杆菌抗原的MAbs的杂交瘤。在六个特异性克隆中，选择产生IgM且与所有56株类鼻疽杆菌分离株反应的克隆5F8进行进一步表征，并评估其可能的诊断潜力。针对各种类鼻疽杆菌抗原的间接ELISA、直接细菌凝集和免疫荧光试验结果表明，5F8与表面包膜反应，可能特异性地与脂多糖的一个表位反应。该抗体可轻松用于在原代培养物或模拟血液培养物中鉴定类鼻疽杆菌。该抗体还用于制备亲和纯化抗原，用于间接ELISA，该方法对检测急性败血症型类鼻疽病患者的循环抗体具有高度敏感性和特异性。
