CAJ | 학술논문

目的：利用原核表达并纯化的幽门螺杆菌（helicobacter pylori ，Hp）OipA6重组蛋白建立间接酶联免疫吸附试验（ELISA）检测方法。方法以纯化的 Hp OipA6重组蛋白作为抗原，包被酶标板，加入血清标本和酶标抗人 IgG，优化反应条件，建立间接 ELISA 检测方法并与 oipA 基因检测结果进行比较。结果重组抗原 OipA6浓度为1.0 mg／ml，包被2～8℃过夜，血清稀释度为1∶100，抗人 IgG 稀释度为1∶4000，二抗孵育时间为40 min，显色时间为15 min 时，建立的间接ELISA 法反应最好。与 oipA 基因检测结果比较，该间接 ELISA 法的敏感度和特异度均＞95％。结论通过包被重组OipA6蛋白建立的间接 ELISA 检测方法可用于高毒株 Hp 感染的辅助诊断。
목적：이용원핵표체병순화적유문라간균（helicobacter pylori ，Hp）OipA6중조단백건립간접매련면역흡부시험（ELISA）검측방법。방법이순화적 Hp OipA6중조단백작위항원，포피매표판，가입혈청표본화매표항인 IgG，우화반응조건，건립간접 ELISA 검측방법병여 oipA 기인검측결과진행비교。결과중조항원 OipA6농도위1.0 mg／ml，포피2～8℃과야，혈청희석도위1∶100，항인 IgG 희석도위1∶4000，이항부육시간위40 min，현색시간위15 min 시，건립적간접ELISA 법반응최호。여 oipA 기인검측결과비교，해간접 ELISA 법적민감도화특이도균＞95％。결론통과포피중조OipA6단백건립적간접 ELISA 검측방법가용우고독주 Hp 감염적보조진단。
Objective To establish an indirect ELISA method with Helicobacter pylori (HP)recombinated antigene OipA6 as coating antigen after prokaryotic expression and purification.Methods The ELISA plate was coated with Hp OipA6 recom-binant protein as antigen.Serum specimens and enzyme labeled anti human IgG were added.The reaction conditions was op-timized.The results of indirect ELISA were compared with the detection of oipA gene.Results When Hp OipA6 recombi-nant protein was 1.0 mg/ml,the condition of coating was 2~8℃ one night,the dilution of serum specimens was 1∶100 and enzyme labeled anti human IgG was 1∶4 000,the incubation time of enzyme labeled anti human IgG was 40 min,the chro-mogenic time was 15 min,the result of ELISA was the best.The sensitivity and specificity of the indirect ELISA were both higher than the detection of oipA gene.Conclusion It suggested that the indirect ELISA,in which Hp OipA6 recombinant protein were coated,may be an appropriate assisted method in detection of high virulent Hp infection.