title: [Identification of genotype III Japanese encephalitis virus isolated in Zhejiang province].
abstract: OBJECTIVE
  To investigate the molecular characteristics and genotype of Japanese encephalitis virus (JEV) found in vector mosquitoes in Zhejiang province from 1982 to 1983.
  
  
  METHODS
  A total of 3188 mosquitoes were collected in Dinghai district and Yiwu city in Zhejiang province, during year 1982 and 1983. The virus was isolated by C6/36 cell, and then identified by hemagglutination inhibition test. The isolated strains were activated in year 2011, and plaque forming unit (PFU) were applied to test the virus titer. The suckling rats were tested under intracranial inoculation, where PrM and E genes were amplified and sequenced. Their nucleotide and amino acid sequences were analyzed and compared with the JEV vaccine strain SA14-14-2 and the JEV isolated in Zhejiang province during 2007 and 2010; and phylogenetic tree were constructed by bioinformatic software.
  
  
  RESULTS
  From the 3188 mosquitoes captured, eleven virus strains were isolated and found to be able to cause cytopathogenic effect (CPE) in C6/36 cells within 72 hours. Virus titer ranged from 2.5 to 6.47 lg PFU/ml. The suckling rats would die within 72 hours since the inoculation. The phylogenetic analysis with the PrM and E genes showed that the JEV isolated in Zhejiang during 1982 and 1983 belonged to genotype III; while the JEV isolated in Zhejiang during 2007 and 2010 belonged to genotype I. The analysis of E genes from 5 isolated strains found that the homology of nucleotide sequence was over 98.9%, and the homology of amino acid sequence was over 99.8%. The compare between the 5 virus strains and the vaccine strain SA14-14-2 found 10 common amino acid variation sites, and showed that the homology of nucleotide sequence was over 97.7%, and the homology of amino acid sequence was 99.2%. The compare between the 5 virus strains and the JEV isolated in Zhejiang during 2007 and 2010 found a 87.7% - 87.9% nucleotide homology and an over 98.8% amino acid homology.
  
  
  CONCLUSIONS
  The JEV isolated from the mosquitoes in Dinghai district and Yiwu city between year 1982 and 1983, were genotype III.
authors.affiliation: Virus Institute, Center for Disease Control and Prevention of Zhejiang Province, Hangzhou 310051, China. jyyan1960@hotmail.com
authors.forename: Ju-ying
authors.initials: JY
authors.lastname: Yan
authors.forename: Jian-yue
authors.initials: JY
authors.lastname: Wang
authors.forename: Zhong-fa
authors.initials: ZF
authors.lastname: Wang
authors.forename: Zhong-wen
authors.initials: ZW
authors.lastname: Yu
authors.forename: Zhong-qing
authors.initials: ZQ
authors.lastname: Yu
authors.forename: Jia-yue
authors.initials: JY
authors.lastname: Zhou
authors.forename: Yan-jun
authors.initials: YJ
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002681:China; D009033:Culicidae / Q000821:virology*; D004664:Encephalitis Virus, Japanese / Q000145:classification* / Q000302:isolation & purification*; D004672:Encephalitis, Japanese / Q000821:virology; D005838:Genotype; D010802:Phylogeny; D051381:Rats