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目的研究猪链球菌(Streptococcus suis,S.suis)锌转运蛋白A(ZnuA)对小鼠接种致死量2型猪链球菌(S.suis 2)菌株的免疫保护作用,为进一步研究S.suis 2亚单位疫苗奠定实验基础。方法采用聚合酶链反应(PCR)检测znuA基因在不同血清型S.suis中的分布情况;蛋白免疫印迹(western blot)检测ZnuA在S.suis 2的表达,利用流式细胞术对ZnuA进行细胞定位,动物实验研究ZnuA蛋白的免疫保护作用。结果除血清型17、21和30型菌株及荷兰分离株T15菌株外,其他30个血清型S.suis菌株、7996菌株以及3株S.suis 2型国内分离菌株基因组中均扩增到目的条带;S.suis 2菌体蛋白与兔抗ZnuA蛋白的血清能发生特异性反应;兔抗ZnuA血清标记S.suis 2的荧光强度明显较高;ZnuA重组蛋白免疫组小鼠死亡率较低。结论 ZnuA是一种具有免疫保护作用的蛋白,可作为S.suis的亚单位疫苗候选分子。
Objective To study the protective effect of Streptococcus suis (S. suis) zinc transporter A (ZnuA) on the lethal dose of S. suis 2 in mice. To study the protective effect of S. suis 2 Subunit vaccines lay the foundation for the experiment. Methods The distribution of znuA gene in different serotypes of S.suis was detected by polymerase chain reaction (PCR). The expression of ZnuA in S.suis 2 was detected by western blot. The expression of zuuA was detected by flow cytometry Positioning, animal experiments to study ZnuA protein immunoprotection. Results In addition to the serotypes 17, 21 and 30 strains and the Netherlands isolates T15 strains, the other 30 serotypes of S. suis, 7996 and 3 strains of S. suis 2 were amplified in the genome ; S.suis 2 cell protein reacted specifically with rabbit anti-ZnuA protein; the fluorescence intensity of anti-ZnuA serum-labeled S.suis 2 was significantly higher; the mortality of ZnuA recombinant protein-immunized mice was lower. Conclusion ZnuA is an immunoprotective protein that can be used as a subunit vaccine candidate for S.suis.