CAJ | 학술논문

目的构建梅毒螺旋体(Tp)膜脂蛋白Tp0821的重组质粒,表达、纯化其相应蛋白,研究其免疫活性.方法构建重组质粒pQE32/Tp082l,诱导表达其相应蛋白,以纯化的重组蛋白免疫新西兰兔,制备多克隆抗体并测定其效价.建立间接ELISA法,检测80份梅毒参比血清、临床经FTA-ABS确诊的阳性血清各150份.结果成功构建重组质粒pQE32/Tp0821,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析重组蛋白分子量与预期结果相符.将亲和层析后所获高纯度重组蛋白免疫新西兰兔,制备的多克隆抗体效价达1:6400.间接ELISA法检测梅毒参比血清、临床梅毒患者血清,与间接免疫荧光螺旋体抗体吸附试验(FTA-ABS)法比较,其灵敏度、特异度分别为92.6%和98.6%,差异有统计学意义(P<0.05).结论 Tp0821重组蛋白具有较好的免疫活性,可用于梅毒血清学检测.
목적 구건매독라선체(Tp)막지단백Tp0821적중조질립,표체、순화기상응단백,연구기면역활성.방법 구건중조질립pQE32/Tp082l,유도표체기상응단백,이순화적중조단백면역신서란토,제비다극륭항체병측정기효개.건립간접ELISA법,검측80빈매독삼비혈청、림상경FTA-ABS학진적양성혈청각150빈.결과 성공구건중조질립pQE32/Tp0821,십이완기류산납-취병희선알응효전영(SDS-PAGE)분석중조단백분자량여예기결과상부.장친화층석후소획고순도중조단백면역신서란토,제비적다극륭항체효개체1:6400.간접ELISA법검측매독삼비혈청、림상매독환자혈청,여간접면역형광라선체항체흡부시험(FTA-ABS)법비교,기령민도、특이도분별위92.6%화98.6%,차이유통계학의의(P<0.05).결론 Tp0821중조단백구유교호적면역활성,가용우매독혈청학검측.
Objective To construct a recombinant plasmid encoding Tp0821,a membrane lipoprotein of T. pallidum,express and purify this protein,and to evaluate its immunocompetence.Methods The recombinant plasmid pQE32/Tp0821 was constructed and induced to express the corresponding protein.Then,New Zealand rabbits were immunized with purified recombinant protein to prepare polycional antibodies,and the titer of polyclonal antibody was determinated.Indirect ELISA was developed with the recombinant protein of T. pallidum as coating antigen to detect 80 control sera and 150 FTA-ABS-positive sera.Results The recombinant plasmid pQE32/Tp0821 was constructed and a fusion protein with expected molecular weight was expressed.Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the antibody titer reached 1:6400.Compared with FTA-ABS test,the indirect ELISA showed a sensitivity and specificity of 92.6%and 98.6%,respectively,in the detection of control and clinical sera.Conclusion The recombinant protein Tp0821 shows excellent immunocompetence,which can be applied to the serological diagnosis of syphilis.