利用间接ELISA定量分析伤寒沙门菌表面呈现表达的流感病毒抗原

Indirect ELISA for Quantitation of Surface Display of Influenzal Antigen in Salmonella enterica serovar Typhi

目的:建立定量检测伤寒沙门菌表面呈现表达的流感病毒抗原的间接ELISA方法。方法:ELISA板以2.5%的戊二醛溶液预处理,将呈现表达M2e等流感病毒抗原表位的伤寒沙门菌的全细胞抗原在ELISA板上进行干燥包被,通过间接ELISA确立全菌抗原的最佳包被浓度;分别采用化学合成多肽M2e和GST-M2e融合蛋白干燥包被ELISA板,绘制标准曲线,对沙门菌表面呈现表达的抗原进行定量分析;对多肽和融合蛋白干燥包被进行比较,同时确立用于定量表面展示量的回归方程。结果:用多肽包被测定的呈现表达的M2e分子数为9.8×104,以GST-M2e包被测定的呈现表达的M2e分子数为1.3×105。结论:利用全菌干燥包被ELISA板可以对伤寒沙门菌表面呈现表达的抗原进行很好的定量分析。

Objective：To develop a indirect ELISA for quantitating surface display of influenzal antigen in Salmonella enterica serovar Typhi.Methods：The whole cell antigen of S.enterica serovar Typhi was integrally dry-coated in ELISA plates which were pretreated by glutaraldehyde of 2.5% concentration.Using the method of indirect ELISA,a hundred million of whole cells per milliliter in buffer were selected for the proper dry-coating concentration.A series concentration of chemical synthetic peptides of M2e and fusion protein of GST-M2e were used to dry-coat in ELISA plates,and draw standard curve,respectively.According to the two standard curves,the surface display expression of influenzal antigen was confirmed,respectively.Results：Indirect ELISA was successfully constructed to quantitate the influenzal antigen,and the surface expression of 9.8×104 and 1.3×105 molecules M2e per bacterium was confirmed,respectively.Conclusion：The whole cell dry-coating could be rather helpful to develop of indirect ELISA quantitating surface display of influenzal antigen in Salmonella enterica serovar Typhi.