RT-PCR Detection and Sequence Analysis of Puumala Virus in Jilin Province

Objective To establish RT-PCR method for Puumala virus(PUUV) detection in order to confirm existence of Puumala virus in Jilin province and analyze gene sequence of Puumala virus.Methods A method based on RT-PCR was used for the detection of Puumala virus in lung samples of Clethrionomys rufocanus trapped in Fusong district of Jilin province.Sequence analysis of nucleocapsid protein coding sequences among S segment of Puumala virus was performed and phylogenetic tree was constructed.Results The research showed the existence of Puumala virus in Fusong district,the positive rate was 7.34% for the samples collected in 2005,and 14.29% in 2006,which showed a increment trend year after year. Sequence analysis showed that the nucleotide homology between Fusong virus and other PUUV was 81.1%-83.3%.Phylogenetic analysis revealed that the Puumala virus found in Fusong was related to the isolates from Japan and Korea,and occupied more ancestral node in the evolution.Conclusion A new subtype of Puumala virus is probably existed in Fusong district of Jilin province.