Study On Pathogentic Detection Result Of HFMD Cases And Whole Genome Feature Of Enterovirus71Isolated In Ningbo，zhejiang Province，china

Objective1、Through to detect the clinical manifestations of hand, foot and mouth disease, Toinvestigate distribution of the pathogen of HFMD in Ningbo，which may be provide reference forpreventing and controlling the HFMD.2、On the base of complete genome sequence, analyse of feature of the EV71completegenome sequence, The Homology and Phylogenetic analysis based on different regions of EV71genome sequence, To predict the secondary structure and B cell epitopes of enterovirus type71ofVP1Gene of C4subgenotype, filtering of positive selection sites.Methods1、446samples were collected from clinically diagnosed cases with HFMD，real-timequantitative RT-PCR or RT-PCR were carried out for nucleinic acid identification and typing.compared and analyzed by using bioinformatics software including DNAMAN and DNAStar,statistic analysis by using software SPSS17.0, entirety reflect the dynamic evolution.2、Stools specimens was collected from clinically diagnosed cases with HFMD in Ningbo，real-time quantitative RT-PCR were carried out for typing，identified EV71was performed forViral isolation and whole genome amplification and sequencing, and another EV71VP1sequencing, compared and analyzed by using bioinformatics software including MEGA4.0.2、DNAStar、DNAMAN、BioEdit、PAML3.15. The Homology and Phylogenetic analysis basedon different regions of EV71genome sequence, To predict the secondary structure and B cellepitopes of enterovirus type71of VP1Gene of C4subgenotype, filtering of positive selection sites.Results1、Among the446clinical samples，346(77.58%)cases were tested positive, including191cases of EV71positive;121cases of CoxA16positive;13cases of CoxA6positive;7cases ofCoxA10positive;4cases of CoxA12and ECHO9positive;2cases of CoxA2positive;1cases ofCoxA4、CoxB1、CoxB4and other enterovirus positive. the proportion of male and female in thecases of HFMD was2.035:1,present peaked in late spring and early summer. The cases in2010 the number of testing less than in2009, EV71was the dominant pathogen of the HFMD in Ningboin2009～2010(P<0.05).2、The full length genome of Ningbo strain (not including Poly A tail)was7406bp, there wasonly one ORF between two UTRs which encoding a polyprotein including2193amino acids.No insertion or deletion was included in the coding region. The Homology and phylogeneticanalysis revealed that Ningbo strains clusterd within the C4a evolution branch of C4subgenotype.The B cell epitopes was probably located at the regions of39~40、159~167、212~220、287~290of VP1Gene. EV71coding areas are not obvious effects of evolution stress, and in purifiedchoose state.Conclusions1、 Pathogentic:446samples were detected from clinically diagnosed cases withHFMD,including191cases of EV71positive;121cases of CoxA16positive;13cases of CoxA6positive;7cases of CoxA10positive;4cases of CoxA12and ECHO9positive;2cases of CoxA2positive;1cases of CoxA4、CoxB1、CoxB4and other enterovirus positive.2、Among the446clinical samples，346(77.58%)cases were tested positive，the proportionof male and female in the cases of HFMD was2.04:1，present peaked in late spring and earlysummer. The cases in2010the number of testing less than in2009.3、Ningbo EV71strain complete genome sequence was7406bp, the GeneBank accessionnumber were JF830007、JN864018~JN8640204.4、Homology analysis: Ningbo EV71strain hared the highest similarity with the C4asubgenotype.5、Phylogenetic analysis: Ningbo strains clusterd within the C4a evolution branch of C4subgenotype.6、Prediction of B cell epitope of enterovirus type71of VP1gene: The B cell epitopes wasprobably located at the regions of39~40、159~167、212~220、287~290of VP1Gene.7、Filtering of positive selection sites: EV71coding areas are not obvious effects ofevolution stress, and in purified choose state.