部分纯化抗原半定量检测幽门螺杆菌感染

Purification of the specific antigens of Helicobacter pylori for semiquantitative serodiagnosis

目的：鉴定并提取幽门螺杆菌（ＨＰ）特异性抗原，建立测定抗ＨＰ抗体的半定量免疫酶技术。方法：１０株ＨＰ可溶性蛋白用于十二烷基磺酸钠－聚丙烯酰胺凝胶电泳和蛋白印迹分析，凝胶层析提取特异性抗原，方阵滴定建立免疫酶方法。纯化抗原与全菌抗原同批检测１３６例患者，以蛋白印迹分析作为金标准。结果：ＨＰ主要特异性抗原条带是６６０００，５９０００，３１０００和２５０００，ＳｅｐｈａｄｅｘＧ－２００柱层析第１峰提取物含其中３种抗原成分。纯化抗原ＥＬＩＳＡ测定抗ＨＰＩｇＧ，敏感性９８．９％，特异性９２．５％，准确性９７．０％，优于应用全菌抗原的检测结果。结论：本实验建立的检测抗ＨＰＩｇＧ方法，具有敏感、特异、可靠的特点。

Objective:To extract and analyse the specific antigens of Helicobacter pylori (HP), which are to be used for establishment of a new immunoenzyme technique for anti HP detection. Methods: The soluble proteins from ten HP strains were analysed with SDS PAGE and Western bloting, and a newly elaborated method with square titration was established for detecting anti HP IgG by ELISA. 136 serum samples were examined simultaneously with the specific antigens and whole cell antigens of HP and immunoblot was used as a gold standard. Results: The major immunogenic polypeptides of HP were the bands at 66 000 ,59 000,31 000 and 25 000, out of which three antigens were obtained for detection. The results obtained by the new immunoenzyme methods were a sensitivity of 98.9%, specificity 92.5%, accuracy 97.0 %, respectively. Conclusion: Depurant antigens used for measuring anti HP IgG are superior to whole cell antigen in ELISA.