标记人血清抗体IgG胶体金免疫法对不同标本幽门螺杆菌抗原的测定  

Detection of Helicobacter pylori antigen with gold-labeled IgG purified from H. pylori infected patients

作　　者：贾克东[1] 丁元军 刘东林[1] 刘晖[1] 罗斌[1] 

机构地区：[1]江西省人民医院,南昌市330006 [2]江西省景德镇市第二人民医院

出　　处：《中华实验和临床感染病杂志（电子版）》2009年第1期1-3,共3页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：江西省卫生厅学术与技术带头人资助项目

摘　　要：目的探讨胶体金免疫法检测患者不同体液中幽门螺杆菌(H.py-lori,HP)抗原的可行性。方法(1)筛选高效价人抗-H.pylori免疫血清,依次应用盐析和层析技术分离纯化获得抗-HP血清IgG;(2)采用柠檬酸三钠还原法制备胶体金颗粒,标记纯化的抗-HP抗体,制成金标抗体;(3)将纯化的特异性人抗-HP抗体固定于硝酸纤维素膜上,制成胶体金标免疫反应模板。分别取粪便、胃液、血清与反应模板结合,然后与金标记的人抗-HP抗体于37℃进行反应,阳性结果呈现肉眼可见的红斑。结果分别以金免疫HP抗原检测、尿素酶实验、血清学抗体检测3种方法对60例患者进行检测对比。HP抗原金免疫检测以胃液准确性最高,为69.04%,粪便检测次之,为57.14%,血清检测准确性最低,为40.48%;检测胃液与传统的尿素酶实验的符合率为61.67%,与血清HP抗体检测的符合率为65%。结论胶体金免疫法测定HP抗原诊断HP感染是可行的,本方法具有操作简便、快速的特点,尤其以胃液和粪便检测效果更好,但该项实验的敏感性和特异性有待进一步提高。Objective To approach the feasibility and accuracy to detect Helicobacter pylori antigen in different samples by colloidal gold immunoassay. Methods （1） The sera with high titer anti-H, pylori antibedy were gathered. IgG was purified and enriched through ammonium sulfate precipitation and protein A resin affinity chromatography. （2） The colloidal gold were prepared with citrate tri-sodium reduction method. Purified human anti-H, pylori IgG was labled with colloidal gold, which named gold-tagged-antibody. （3） Purified human anti-H, pylori IgG was fixed on the membrane of nitrocellulose. Samples of gastric juice, feces and sera were added on the membrane where antigen-antibody combined,, respectively. The result was shown by gold-tagged-antibody and with red spot on the nitrocellulose membrane when considered as positive sample. Results The samples of gastric juice, feces and serum of 60 patients were detected for H. pylori antigen by H. pylori antigen immune-gold test, the accuracy of gastric juice is 69.04%, 57.14% of feces and 40.48% of serum. The consistent rates between this method and routine urease test and serologic antibody test are 61.67% and 65%, respectively. Condusions It is feasible to detect H. pylori infection by collidal gold immunoassay through anti-H, pylori IgG obtained from sera of H. pylori infected patients, but the sensitivity and specificity need to be improved in the future.

关 键 词：幽门螺杆菌 金标免疫法 抗原诊断 

分 类 号：R446.6[医药卫生—诊断学]