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AIM:TO examine the serological response of patients withupper gastrointestinal diseases and Helicobocter pylori(H pylon)infection to two Hpyloriouter membrane proteins(OMPs)(M,18 000 and M,26 000)acquired by gene recombinanttechnique,and to determine the diagnostic significance ofserological tests derived from these OMPs.METHODS:Recombinant vectors encoding the two HpyloriOMPs were used to transform and express in BL21(DE3)E.COli.After purification with Ni~(2+)-NTA agarose resin,colloidgold kits were prepared with purified recombinant proteinsto detect H pylori infectJon and H pylori-associated diseasesby the immunity-marker technology.We selected 150 patientswith Hpyloriinfection and digestive symptoms without previoustreatment,induding chronic gastritis(n=60),duodenal ulcer(n=30),gastric ulcer(n=30),and gastric cancer(n=30).As controls,33 Hpylori-negative healthy volunteers werealso recruited.Serum samples were collected from allsubjects,and the antibodies to specific proteins of H pyloriwere tested with the colloid gold test kits.The sensitivity,specificity and accuracy of the colloid gold tests wereevaluated,by using the combination of standard diagnosticmethods(~(13)C urea breath test and bacteria culture)andclassic enzyme-linked immunosorbent assay(ELISA)asreference.RESULTS:After purification with Ni~(2+)-NTA agarose resin,the purity of recombinant fusion proteins was about 95%.The recombinant fusion proteins were recognized by thespecific monodonal antibodies against the two Hpylori OMPs,as demonstrated by the ELISA.Of the 150 serum samplesfrom patients infected with Hpylori 141(94.0%)respondedpositively to the recombinant protein with M_126 000,whilethe seropositive rates were 95.0%,96.7%,96.7% and 90.0%for patients with H pylori-associated chronic gastritis,duodenal ulcer,gastric ulcer,and gastric cancer respectively.The sensitivity,specificity,and accuracy of the colloid goldkit with M_1 26 000 protein were 94.0%,97.0%,and 94.5%,respectively.Compared with the classic ELISA,bacteriaculture and ~(13)C urea breath test results in detecting Hpylori- infection,there was no significant difference(P>0.05).Forthe colloid gold kit with M,18 000,the seropositive rateswere 52.0%,40.0%,40.0%,53.3% and 86.7%,respectively,in Hpylori-infected patients,and those with Hpylori-assodatedchronic gastritis,duodenal ulcer,gastric ulcer,and gastriccancer.There was a significant difference(P<0.05)inseropositivity between patient with gastdc cancer(86.7%)and those with other diseases(43.3%).CONCLUSION:The two colloid gold kits derived from therecombinant OMPs are useful tools either for detectingHpyloriinfection,or for,predicting Hpylori-assodated gastricmalignancy.
AIM: TO examine the serological response of patients with upper gastrointestinal diseases and Helicobocter pylori (H pylon) infection to two Hpyloriouter membrane proteins (OMPs) (M, 18 000 and M, 26 000) acquired by gene recombinant technology, and to determine the diagnostic significance of serological tests derived from these OMPs. METHODS: Recombinant vectors encoding the two Hpylori OMPs were used to transform and express in BL21 (DE3) E. COli.After purification with Ni ~ (2 +) - NTA agarose resin, colloidgold kits were prepared with purified recombinant proteinsto detect H pylori infect Jon and H pylori-associated diseases by the immunity-marker technology. We selected 150 patients with Hpylori infection and digestive symptoms without previou treatment, induding chronic gastritis (n = 60), duodenal ulcer (n = 30), gastric ulcer = 30), and gastric cancer (n = 30). As controls, 33 Hpylori-negative healthy volunteers were collected. Serum samples were collected from allsubjects, and the antibodies to specific proteins of Hp yloriwere tested with the colloid gold test kits. sensitivity, specificity and accuracy of the colloid gold tests wereevaluated, by using the combination of standard diagnostic methods (~ (13) C urea breath test and bacteria culture) and classic enzyme-linked immunosorbent assay ) asreference.RESULTS: After purification with Ni ~ (2 +) - NTA agarose resin, the purity of recombinant fusion proteins was about 95%. the recombinant fusion proteins were recognized by the specific monodonal antibodies against the two Hpylori OMPs, as demonstrated by the ELISA.Of the 150 serum samples from patients infected with Hpylori 141 (94.0%) respondedpositively to the recombinant protein with M_126 000, while the seropositive rates were 95.0%, 96.7%, 96.7% and 90.0% for patients with H pylori- associated chronic gastritis, duodenal ulcer, gastric ulcer, and gastric cancer respectively. The sensitivity, specificity, and accuracy of the colloid gold kit with M_1 26 000 protein were 94.0%, 97.0%, and 94.5%, respectively. Compared withthe classic ELISA, bacteriaculture and ~ (13) C urea breath test results in detecting Hpylori-infection, there was no significant difference (P> 0.05) .Forthe colloid gold kit with M, 18 000, the seropositive rateswere 52.0%, 40.0% , 40.0%, 53.3% and 86.7%, respectively, in Hpylori-infected patients, and those with Hpylori-assodated chronic gastritis, duodenal ulcer, gastric ulcer, and gastriccancer.There was a significant difference (P <0.05) inseropositivity between patient with gastdc CONCLUSION: The two colloid gold kits derived from therecombinant OMPs are useful tools either forHpylori infection, or for, predicting Hpylori-assodated gastric malignancy.