检测血清幽门螺杆菌细胞空泡毒素抗体的间接ELISA法初步建立

Preliminary establishment of an indirect ELISA assay to detect IgG of VacA of Hp

目的:以重组细胞空泡毒素(VacA)蛋白为抗原初步建立检测血清VacA抗体间接ELISA法,为制备检测Hp毒株感染的试剂盒奠定基础。方法:Ni-NTA2+树脂纯化重组VacA蛋白,用重组蛋白免疫兔,HelicoBlot试剂盒检测兔血清VacA抗体以鉴定其免疫原性。Westernblot检测其抗原性。并以纯化的重组蛋白为抗原,建立检测血清VacA抗体间接ELISA法,以HelicoBlot试剂盒为标准,确定该方法的特异性与敏感性。结果:以重组蛋白为抗原建立的检测血清VacA抗体的间接ELISA法,敏感性与特异性分别是93.1%和92.5%。结论:以重组蛋白为抗原初步建立检测血清VacA抗体的间接ELISA法,敏感性、特异性高,为制备商品化的试剂盒奠定了基础。

Objective To establish an indirect ELISA with the recombinant protein,which would provide a solid basis for obtaining diagnostic reagent kit detecting Hp toxicity strain infection. Methods The recombinant protein purified with Ni2+-NTA agarose was used to immunize rabbits to prepare antiserum, in which the anti-VacA was detected by helico-blot kit, and its antigenicity was confirmed by Western blot. The indirect ELISA was established with the recombinant protein to detect the IgG in sera of patients infected with Hp, which was compared with helico-blot kit, and then its sensitivity and specifity was evaluated. Results The indirect ELISA based on recombinant protein was successfully established to assay the anti-VacA in serum, and its sensitivity and specificity was 93.1% and 92.5% respectively. Conclusion The indirect ELISA was created to assay anti-VacA in serum provided a basis for the preparation of diagnostic reagent kit detecting Hp toxicity strain infection.