幽门螺杆菌外膜蛋白对Helicobacter pylori感染的诊断  

Helicobacter pylori outer membrane proteins in diagnosis of Helicobacter pylori infection

作　　者：姜政[1] 黄爱龙[2] 郑健[2] 陶小红[1] 蒲丹 王丕龙[1] 

机构地区：[1]重庆医科大学第一附属医院消化科,重庆市400016 [2]重庆医科大学肝炎研究所,重庆市400010

出　　处：《世界华人消化杂志》2004年第7期1588-1592,共5页World Chinese Journal of Digestology

基　　金：重庆市科委应用基础研究计划项目;No.[2002]18-86国家自然科学基金资助;No.30371318~~

摘　　要：目的:通过检测伴有H pylori感染的上消化道疾病患者血清对H pylori OMP的反应性,来探讨基因重组H pylori OMP对H pylori感染及其相关性疾病的诊断价值,为进一步开发诊断试剂盒及其临床应用奠定基础. 方法:分别将已鉴定、测序的含H pylori Mr为18 000, 26 000 OMP重组质粒转化表达菌-大肠杆菌BL21中, 让其大量表达、纯化,制备金标免疫检测试纸;选择2002-01/2002-12因消化道症状来我院就诊的H pylori 阳性患者150例,经胃镜证实:慢性浅表性胃炎60例: 胃溃疡30例;十二指肠球部溃疡30例;胃癌30例,以及33例H pylori阴性的健康者作为对照,分别采用以H pylori Mr为18 000,26 000 OMP为抗原制备金标免疫检测试纸,对183例受试者血清进行特异性抗体检测. 结果:重组蛋白经Ni2+ -NTA琼脂糖树脂纯化后,其纯度高达95%以上,经检测其抗原性良好.用金标免疫试纸对H pylori感染的上消化道疾病患者150例,以及H pylori阴性的健康者33名进行了检测,其结果为: 26 000 OMP对H pylori感染的检出率为94.0%,对慢性浅表性胃炎、胃溃疡、十二指肠球部溃疡以及胃癌患者中H pylori感染的检出率分别为95.0%,96.7%, 96.7%和90.0%,与常规ELISA方法检测结果相比, 二者无显著性差异(X2检验,P>0.05),其敏感性、特异性和准确性分别为94.0%、97.0%、94.5%;而18 000 OMP对H pylori感染患者的检出率为52.0%,对慢性浅表性胃炎、胃溃疡、十二指肠球部溃疡和胃癌患者中H pylori感染的检出率分别为40.0%、40.0%、53.3% 和86.7%,对胃癌患者中H pylori感染检出率86.7%与慢性浅表性胃炎、胃溃疡、十二指肠球部溃疡总检出率43.3%相比,具有显著的差异性(P<0.05).因此以26 000 OMP的金标免疫检测试纸可以作为H pylori染的常规检测方法,他与常规ELISA检测方法相比具有可靠的特异性以及敏感性,同时与18 000 OMP金标免疫检测试纸一起,对胃癌患者中H pylori感染检出率高对胃肠道肿瘤�AIM: To examine the serological response of patient with various upper gastrointestinal diseases and Helicobacter pylori (H pylori) infection to H pylori outer membrane proteins (OMP) with Mr18 000 and 26 000 acquired by gene recombinant technique, and to determine its diagnostic significance. METHODS: The recombinant vectors encoding OMP of H pylori with Mr18 000 and 26 000 identified by restriction enzyme or PCR were used to transform and express in BL21 (DE3) E. coli respectively. After purification with Ni2+-NTA agarose resin, the colloid gold kits were prepared with purified recombinant proteins to detect H pylori infection and disease associated with H pylori by immunity-marker technology. We selected 150 patients with H pylori infection and digestive symptoms without previous treatment, including chronic superficial gastritis (n =60), duodenal ulcer (n =30), gastric ulcer (n =30), and gastric cancer (n =30) during one year. Simultaneously, 33 cases without digestive symptoms and H pylori infection were used as controls. All sera were collected and the antibody responded to specific proteins of H pylori was tested with the colloid gold test kit. Taking as reference a combination of standard diagnostic methods (13C urea breath test, cultivate) and the classic enzyme-linked immunosorbent assay (ELISA) serological tests was com- pared with the results of this technique, and the sensitivity, specificity and accuracy of the colloid gold test were evaluated. RESULTS: After purification with Ni2+-NTA agarose resin, the purities of recombinant fusion proteins were all about 95%. The ELISA results showed that recombinant fusion proteins could be recognized by monoclonal antibody of anti-H pylori with Mr18 000 and 26 000 respectively. The results detected using colloid gold kits were as follows: all patients' sera infected with H pylori showed response to recombinant protein with Mr26 000 were 94.0%, while 95.0%, 96.7%, 96.7% and 90.0% of patients with H pylori-infected chronic superficial gastritis, duodenal ulcer,

关 键 词：幽门螺杆菌 外膜蛋白 Helicobacterpylori感染 诊断 

分 类 号：R57[医药卫生—消化系统]