Abstract：Objective To explore whether the 5′-untranslated region (5′-UTR) and VP1 region ofenterovirus 71(EV71) isolated from severe hand, foot and mouth disease (HFMD) contain potential virulence sites. Methods EV71 strains isolated from fecal specimens collected from HFMD patients with different clinical symptoms were cultured by RD cells. The viral total RNA was extracted. The 5’-UTR and VP1 in genomes were amplified by RT-PCR and then sequenced. The nucleotide sequences of 5′-UTR and VP1 and the amino acid sequences of VP1 were compared, and the homology analysis was performed. The phylogenetic tree was constructed based on the VP1 gene sequences and the 5′-UTR of EV71 with different clinical symptoms. Results A total of 9 EV71 strains (5 strains from severe cases and 4 strains from mild cases) were obtained in this study. The nucleotide homologies of 5’-UTR and VP1 regions were both 94.5%-99.7%, and the amino acid homology of VP1 region was 97.3%-99.9%. There were 67 mutation sites in 5′-UTR of the 9 EV71 strains. Compared with the 4 EV71 strains isolated from mild cases, 2 amino acids (S283 T, A289T) of VP1 from the 5 EV71 strains isolated from severe cases were substituted, and a total of 37 nucleotide positions were mutated in 5′-UTR sequences. The phylogenetic tree based on VP1 sequences showed that the 9 EV71 strains belonged to sub-genogroup C4a, and the strains isolated from different clinical manifestation cases of two groups were clustered in the same branch. Phylogenetic tree based on the nucleotide sequences of 5′-UTR showed that EV71 strains from different clinical manifestation cases exhibited a staggered distribution, and the strains from same clinical manifestation cases did not cluster separately. Conclusion The nine EV71 strains belonged to the sub-genogroup C4a. Nucleotide mutations at 5′-UTR and amino acid substitutions at VP1 may affect the virulence of EV71. Accordingly, further study on the whole genomecharacterization of the EV71 strains and the interaction mechanism between the EV71 strains and the host protein factors are necessary.
张凤娟, 霍锡元, 丁彦红, 李琳, 刘国军, 聂清. 潍坊市9例重症与轻症手足口病患者肠道病毒71型5′非编码区、VP1基因特征分析[J]. 实用预防医学, 2022, 29(3): 265-268.
ZHANG Feng-juan, HUO Xi-yuan, DING Yan-hong, LI Lin, LIU Guo-jun, NIE Qin. Gene characteristics of 5′-UTR and VP1 of enterovirus 71 isolated from 9 patients with severe or mild hand, foot and mouth disease in Weifang City. , 2022, 29(3): 265-268.
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