OBJECTIVE: To isolate Japanese Encephalitis virus (JEV) from mosquitoes collected in Jinan city, Shandong province, and to analyse the genotype of new isolated JEV strains and the characteristic of E gene and PrM gene. METHODS: Isolate virus from mosquitoes collected in 2008 and identify the isolated virus by biological, serological and molecular biological method. PrM and E segments of new isolated JEV were amplified by RT-PCR, the PCR products were purified and sequencing. Multiple alignment, phylogenetic and amino acid (AA) analysis were carried out by Clustal X (1.83) program, MEGA 3.1 and GENEDOS (3.2). RESULTS: Total 15,700 mosquitoes were collected including Culex, Armigeres, Aedes, Anopheline. 20 loads were taken at random, one virus strain was isolated from Culex tritaeriorhynchus and identified to be genotype 1 JEV. The homologue of nucleotide and amino acid of E gene between the new isolated JEV and live attenuated vaccine strain SA14-14-2 were 88.0%, 97.2% respectively. There were 14 amino acid differences in E gene between them. CONCLUSION: One strain of JEV belonging to genotype 1 was isolated firstly and identified in Jinan city, Shandong province.
OBJECTIVE: To isolate Japanese Encephalitis virus (JEV) from mosquitoes collected in Jinan city, Shandong province, and to analyse the genotype of new isolated JEV strains and the characteristic of E gene and PrM gene. METHODS: Isolate virus from mosquitoes collected in 2008 and identify the isolated virus by biological, serological and molecular biological method. PrM and E segments of new isolated JEV were amplified by RT-PCR, the PCR products were purified and sequencing. Multiple alignment, phylogenetic and amino acid (AA) analysis were carried out by Clustal X (1.83) program, MEGA 3.1 and GENEDOS (3.2). RESULTS: Total 15,700 mosquitoes were collected including Culex, Armigeres, Aedes, Anopheline. 20 loads were taken at random, one virus strain was isolated from Culex tritaeriorhynchus and identified to be genotype 1 JEV. The homologue of nucleotide and amino acid of E gene between the new isolated JEV and live attenuated vaccine strain SA14-14-2 were 88.0%, 97.2% respectively. There were 14 amino acid differences in E gene between them. CONCLUSION: One strain of JEV belonging to genotype 1 was isolated firstly and identified in Jinan city, Shandong province.