Sequence And Molecular Epidemiology Analyses Of Hemorrhagic Fever With Renal Syndrome Virus

Objectives To amplify M segments by RT-PCR from Hantavirus (HV) infected rodents in Shandong Provience and sequence them, to understand the genotype of HV and the genetic distances between the M segments by phylogenetic analysis and homology analysis, to describe the epidemic characteristic of HFRS in Shandong Provience and get the HV representative strain, to construct the coloning vector of the whole M gene of the representative strain and sequence it, to analyze the molecular epidemiological characteristics of the representative strain, to facilitate study on the relationship between gene and protein, and to lay foundation for HV subunit vaccine researches.Methods HV antigens in the collected samples were detected by the indirect immunofluorescence assay (IFA). The general and the type primers for SEOV and HTNV were designed by software Primer5. The nucleotide sequences of the partial M (300 bp) segments were amplified by reverse transcription polymerase chain reaction (RT-PCR) from the HV positive samples. The results were compared with the previous strains of Shandong Province in homology and phylogenetics. The HV representative strain of Shandong Provience was got after the gene difference analyses. Coloning vector of the M gene of the HV was constructed and sequenced. Thededuced amino acid was analyzed by protein software to get its biologic characteristics.Results 1. There were 2073 rodents were captured, which have 103 HV positive samples, the infection rate being 4.97%.2. There were 26 M segments (2003~2302nt) were amplified by RT-PCR from positive samples.3. Nucleotide sequence homology analysis showed that they were all SEO types of HV. The nucleotide sequence homology of these samples was from 97.3% to 100.0% and nucleotide sequence homology with other Shandong strains was from 95.5% to 100.0%. The phylogenetic tree analysis show that they all belong to the same SEO subtype S3.4. Cloning vector of the M gene of the Shandong HV representative strain GM04-38 was constructed. The nucleotide sequence homology analysis showed that it was close to strain Z37, the nucleotide sequence homology was 97.3%, and the deduced amino acid homology was 99.2%.Conclusions The M segment of different places in Shandong Provience were amplified successfully. They mainly were SEO type and highly conserved. The representative strain GM04-38 was gotten by the homology analysis and phylogenetic tree analysis. The clone of HV GM04-38 strain M gene was constructed successfully. The analysis of nucleotide and amino acid sequence provided effective data for the study on the mutation principles of HV and the inspection and control of HV at the molecular level. The cloning of the M gene can serve as the foundations for studying the relationship between M gene structures and functions, and to develop the subunit vaccines.