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目的对5株EV71分离株(LCH01、LCH02、BJ01、BJ02、AH01)进行毒力、免疫原性和基因组的比较分析。方法分别用5株EV71分离株攻击Balb/c乳鼠,比较毒力差异;免疫6～8周龄Balb/c小鼠后收集血清,ELISA检测抗体效价,微量中和实验检测中和抗体效价;反转录获得5株EV71的全长cDNA进行全基因组测序;最后,用CLC Main Workbench 5.5软件对序列进行比较分析。结果通过对VP1基因的比较分析发现,这5株EV71分离株均为C4a亚型。1周龄Balb/c小鼠经脑部注射LCH01株,表现出后肢瘫痪和后继死亡;而注射其它4株EV71的小鼠在观察了14 d之后仍然存活。5株EV71血清抗体效价为210～211,中和抗体效价为27～28,无显著差异。LCH01毒株与其它几株序列相比,在5′-NTR、3′-NTR、3D等处均有明显差异。结论这5株EV71分离株为近几年流行的C4a亚型,具有较高的免疫原性;LCH01毒株的毒力高于其它几株,从基因组水平上找到了差异序列,这为EV71疫苗评价和致病机制研究提供了实验数据和实验材料。
Objective To compare the virulence, immunogenicity and genome of five strains of EV71 (LCH01, LCH02, BJ01, BJ02, AH01). Methods Balb / c suckling mice were challenged with five EV71 isolates respectively, and the virulence differences were compared. Balb / c mice of 6-8 weeks old were harvested and the antibody titer was detected by ELISA. Neutralizing antibody titer The full-length cDNA of 5 EV71 strains was reverse transcribed for genome-wide sequencing. Finally, the sequences were analyzed by CLC Main Workbench 5.5 software. Results The comparative analysis of VP1 gene revealed that all five EV71 isolates were C4a subtypes. One week old Balb / c mice were injected intracerebrally with LCH01 strain, showing hindlimb paralysis and subsequent death; mice injected with the other four EV71 survived after 14 days of observation. Five strains of EV71 serum antibody titer of 210 to 211, neutralizing antibody titer of 27 to 28, no significant difference. Compared with several other sequences, the LCH01 strain showed obvious differences in 5'-NTR, 3'-NTR and 3D. Conclusion These five EV71 isolates are highly sub-genotypes of C4a that have been popular in recent years. The virulence of LCH01 strain is higher than that of other strains, and the difference is found at the genomic level. This is EV71 vaccine Evaluation and pathogenesis studies provided experimental data and experimental material.