Objective To analyze the conding region of hantanvirus S gene and predict the structure of nucleoprotein for diagnostic antigen study.Methods RT-PCR was used to amplify the S gene of hantanvirus Hunan03 strain after designing specific primers.The amplification product was cloned into pGM-T vector and then the recombinant vector was transformed into E.coli TOP10,gene sequencing was carried out after blue-white selection and PCR screening for positive clones.The database of NCBI and Swiss-Prot/TrEMBL were used to predict and analyze the structure,biological characteristics and protein structures of S gene.Results The amplification product was about 1290 bp,the pGM-T/S vector was constructed and successfully sequenced,the whole length of the open reading frame (ORF) was composed of 1290 nucleotide residues,among them the GC content was 44.11％ and the AT content was 55.89％,it was composed of 429 amino acids (20 kinds),the accession number of the sequence submitted to GenBank was JN712306,its homology of nucleotides to the 76-118 strain was 83％ and the homology of amino acids was 98％,ten nonspecific variation sites were found.The grand average of hydropathicity was-0.405.There were three transmembrane domains and four non transmembrane domains in the secondary structure of nucleoprotein including 55％ of helix structure,6.1％ of sheet structure and 38.9％ of loop structure.Conclusion The bioinformatics analysis of Hunan03 strain S gene might be important for provide the substructure data to reveal the significance of S gene characteristics on hemorrhagic fever renal syndrome (HFRS) prevention and control.

To investigate the genotype and characteristic of hantavirus M and S hemorrhagic fever with renal syndrome(HFRS),lung tissues of rats were screened by immunofluorescent antibody assay(IFA),and the total RNA was extracted from the serum of HFRS patients and IFA positive lung tissues.RT-PCR was applied to amplify the M and S gene of hantavirus after specific primers were designed.PCR positive product was recovered and purified before cloning to the pMD-18 T vector and the sequencing was carried by ABI3730 gene sequencer.The obtained sequencings were compared to the HTNV and SEOV sequences of hantavirus for nucleotide homology analysis and applied MEGA5.0.4 software to construct the phylogenetic tree.The yellow-green needle-like particles were scattered in the positive lung tissues under fluorescence microscope.The PCR products of M and S gene was about 490bp and 593bp with general primers and the further amplification with HTNV typing primers to the M and S gene was respectively 242bp and 276bp,while the amplified results of M and S gene were negative with the primers of SEOV.The sequences of Hunan01,Hunan02 and Hunan03 had a high nucleotide homology to the 84FLi and SN7 strains,which had a lower homology to the Gou3 strain.Hunan01,Hunan02 and Hunan03 were belonged to H5 subtype of HTNV.In conclusion,the technology of genotyping is appropriate for hantavirus diagnosis,and the HTNV patients and animal hosts are exist in Hunan Province.