抗角蛋白抗体免疫印迹检测技术的建立及临床应用

Clinical Application and Establishment of the Immune-blotting Method of Detecting AKA

目的建立抗角蛋白抗体(Anti-keratin antibody,AKA)的免疫印迹检测技术,探讨其临床适用性和实用性。方法提取大鼠食管上皮组织的角质层抗原,通过SDS-PAGE电泳后将其转移到硝酸纤维素(NC)膜上,对病人标本进行AKA的免疫印迹检测,以间接免疫荧光法作对照。结果类风湿性关节炎(RA),系统性红斑狼疮(SLE)和干燥综合征(SS)的AKA阳性率分别为45.8%,8.6%和16.7%。结论免疫印迹法检测AKA比间接免疫荧光法检测具有更好的灵敏度,而且操作简便,结果易读。

Objective To establish an immune-blotting method of detecting anti-keratin antibody（AKA）and discuss its applicability and practicability in clinic. Methods The antigen of keratin was extracted from rat＇s esophagus epithelium tissue, and then transferred on the nitric acid cellulose （NC） membrane after having electrophoresed through SDS-PAGE glue. It was used to detect AKA in the serum of patients, meanwhile contrasted with the indirect immunofluorescence method. Results The positive rates of detecting AKA were 45.8%,8.6% and 16.7% in RA,SLE and SS patients respectively. Conclusion The immune-blotting method of detecting AKA is the same sensitivity to the indirect immunofluorescence method,and it is an easy and simple way in operating and reading result.