Genotyping And DNA Sequence Analysis Of Hantaviruses

Object: To investigate the hantavirus genotype carried by Apodemus agrarius and A.peninsulae in Heilongjiang Province,Jilin Province and Iner Mongolia Autonomous Region, Establishing the methods of amplifing the full-S fragment to affirm genetic subtypes and its evolution.Clarifying the characteristics of Hantavirus, Providing scientific basis.for prevention and control hemorrhagic fever with renal syndrome; To investigate the hantavirus genotype and features carried by Wistar rats, local R.norvegicus and hemorrhagic fever with renal syndrome patients in Shenyang. Establishing the methods of amplifing the full-M fragment, providing strong evidence.for analyzing of Hantavirus and the host.Methods:Rodents were captured in the main epidemic areas.Hantavirus antigens and IgM , IgG antibodies in rats lungs were detected by indirect immunofluorescnce assay(IFA). Partial and complete M and S segments were amplified in antigen-positive samples by RT-PCR, and then sequenced and analyzed.Hantavirus was isolated from Vero E6.Results: 11 positive samples were identified in A. agrarius, A. peninsulae in Heilongjiang Province,Jilin Province and Iner Mongolia Autonomous Region. The genetic analysis of the complete S segments indicated that the subtype of hantaan virus belong to HTNV6 and 7. Establishing the RT-PCR methods of amplifing the full-S fragment. Virus strains that HAa174 was isolated success from Vero E6.57 rates were collected in the main epidemic areas of Shenyang, Hantavirus specific-antigens were identified in 26 out of the 57 Wistar, 27 epidemic hemorrhagic fever patients, 15 positive samples of R.norvegicus.The genetic analysis of the partial and complete S and M segments indicated that hantavirus-positive samples were Seoul virus in Shenyang. The further genetic and phylogenetic analysis indicated that hantavirus-positive samples were grouped into 2 genetic subtypes, SEOV1 and SEOV3. Establishing the methods of amplifing the full-M fragment. Virus strains that Wistar 41 was isolated success from Vero E6.Conclusion: The results indicated that the virus carried by A.agrarius and A.peninsulae in Northeast of China was HTNV, The subtype of HTNV were HTNV6 and HTNV7. And the main genetic subtype of Hantavirus is S1 and S3 of Seoul virus in Shenyang.