Sequence and antigenicity analysis for VP4s of EV71 strains isolated from Children in Beijing / 病毒学报

ABSTRACT

In order to learn about the correlation between the sequences of VP4 of EV71 and clinical symptoms of patients and analyze the antigenicity of VP4 of EV71, as well as the cross-reactivity with VP4 of CA16, the sequences of VP4 gene from 10 EV71 strains isolated from infants and children with hand, foot and mouth diseases (HFMD) during 2007 to 2009 were determined through standard molecular cloning protocols, and the results were analyzed by EditSeq and MegAlign of DNAStar. Full-length genes of VP4s of EV71 and CA16 were amplified from virus isolates and expressed in E. coli. Then the expressed VP4s were used as antigens to detect IgG antibody in 189 sera samples from people taking health check up and patients of non-HFMD by Western-Blot. They were also used to detect IgM antibody in 14 of sera samples from infants and children with EV71 infection and 12 of sera samples from those with CA16 infection. The nucleotides identities among these 10 sequences of VP4s isolated in our lab were 94.20% - 100.00% and the deduced amino acids were identical. There was no consistent divergence between the sequences of serious cases and those from general HFMD cases. Phylogenetic analysis based on VP4s indicated that these 10 VP4s of EV71 belonged to C4. The nucleotide identities between EV71 VP4 (s67) and CA16 VP4 (s401) was 69.60% and the deduced amino acids identities was 78.60%. In the detection of IgG, the sera-positive rate for EV71 VP4 was 38.10% and the sera-positive rate of CA16 VP4 was 58.20%. The difference in the sera-positive rate between them was significant (chi2 = 15.30, P < 0.01), suggesting that the expressed VP4s of EV71 and CA16 were of good antigenicity and not cross-reactive. There was no positive reaction detected for IgM against VP4s for EV71 or CA16. The data from this study reveal important information for the further study of EV71.