Faulde M, Cremer J, Zöller L
Department of Medical Microbiology, Ernst-Rodenwaldt-Institute, Koblenz, Germany.
Electrophoresis. 1993 Sep;14(9):945-51. doi: 10.1002/elps.11501401150.
An immunoblot method has been evaluated to diagnose Helicobacter pylori infection serologically by comparing 69 serum specimens from patients with a positive Gram stain and/or culture result and a positive urease test on biopsy material, as well as 51 serum specimens from patients with at least 4 negative urease tests, and negative microscopy and culture results. Sensitivity and specificity was found to be 100%. Recognition of the cross-reacting flagellin (66 kDa), flagellar sheath protein (51 kDa), and a 14 kDa protein are not a criterion for a current H. pylori infection. On the other hand, any combination of at least two of the 180, 120, 90, 75, 67, 29.5 and 19 kDa bands were diagnostic of infection. Three H. pylori strains, which were compared with both gel electrophoretic analyses and immunoblot reactivity, exhibited in part strong qualitative and quantitative differences that particularly affect the 120 kDa pathogenic factor, the large urease subunit and other proteins especially in the molecular mass range from 50 to 67 kDa. IgG immunoblot patterns showed that the choice of H. pylori strain, as well as a reproducible and standardizable antigen preparation, is of great importance for the reliability of serodiagnostic tests. The immunoblot method was found to be a valuable tool for the semi-quantitative confirmation of results achieved with other serological methods as well as optimization and quality control of the antigens used for serodiagnostic purposes.
已对一种免疫印迹法进行评估,通过比较69份来自革兰氏染色和/或培养结果呈阳性且活检材料尿素酶试验呈阳性的患者的血清标本,以及51份来自至少4次尿素酶试验呈阴性、显微镜检查和培养结果均为阴性的患者的血清标本,来进行幽门螺杆菌感染的血清学诊断。结果发现其敏感性和特异性均为100%。识别交叉反应鞭毛蛋白(66 kDa)、鞭毛鞘蛋白(51 kDa)和一种14 kDa蛋白并非当前幽门螺杆菌感染的标准。另一方面,180、120、90、75、67、29.5和19 kDa条带中至少两条的任何组合均可诊断感染。对三株幽门螺杆菌菌株进行了凝胶电泳分析和免疫印迹反应性比较,结果显示它们在部分定性和定量上存在显著差异,尤其影响120 kDa致病因子、大尿素酶亚基以及其他蛋白质,特别是分子量在50至67 kDa范围内的蛋白质。IgG免疫印迹模式表明,幽门螺杆菌菌株的选择以及可重复和标准化的抗原制备对于血清学诊断试验的可靠性至关重要。免疫印迹法被认为是一种有价值的工具,可用于半定量确认其他血清学方法所获得的结果,以及用于血清学诊断目的的抗原的优化和质量控制。