细粒棘球蚴特异性诊断抗原Eg-07279的制备及免疫原性研究

目的 筛选出包虫病特异性诊断抗原分子并验证其免疫原性。方法 对国家人类基因组南方研究中心公布的细粒棘球绦虫测序数据进行分析,利用生物信息学的方法筛选出六钩蚴中不表达且原头蚴中高表达的抗原分子Eg-07279,经重组克隆、表达后用亲和层析法纯化获得重组蛋白rEg-07279;重组蛋白免疫小鼠检测其特异性 IgG 水平并使用Western blot 验证其免疫原性。结果 筛选出的抗原分子Eg-07279经克隆、表达、纯化获得重组蛋白。利用该重组蛋白免疫小鼠,ELISA检测结果显示Eg-07279产生的特异性IgG水平(2.559 ± 0.125)明显高于空白组(0.319 0±0.01),差异有统计学意义(P<0.01);Western blot检测结果显示原头蚴继发感染组和重组蛋白免疫组血清均可识别该重组蛋白而空白对照组鼠血清不能识别。结论 获得细粒棘球蚴差异表达抗原Eg-07279并证实该重组蛋白具有较好的免疫原性,是较好的诊断抗原候选分子。

Preparetion and analysis on the immunogenicity of Echinococcus granulosus-specific diagnostic antigen Eg-07279

We screened echinococcosis-specific diagnostic antigen and verified its immunogenicity. The sequencing data of Echinococcus granulosus released by the National Human Genome Research Center was analyzed. The bioinformatics method was used to screen out Eg-07279 antigen gene Eg-07279, which was not expressed in Echinococcus granulosus and highly expressed in the original metacercariae. After expression, the recombinant protein rEg-07279 was purified by affinity chromatography. The specific IgG level was detected in the recombinant protein immunized mice and the immunogenicity was verified by Western blot. The selected antigen molecule Eg-07279 was cloned, expressed and purified to obtain the recombinant protein. The results of ELISA showed that the specific IgG level of Eg-07279 (2.559±0.125) was significantly higher than that of the blank group (0.319 0±0.01), the difference was statistically significant (P<0.01); the Western blot results showed that the recombinant plasmids could be recognized by the secondary infection of the protoscolex and the serum of the recombinant protein immunized group, while the serum of the blank control group was not recognized by the serum. In summary, the Eg-07279 antigen of E. granulosus was obtained and it was proved that the recombinant protein has good immunogenicity and is a good diagnostic antigen candidate molecule.