Molecular epidemiology of Japanese encephalitis viruses isolated in Yunnan province, 1977-2010

Abstract

Objective: To understand the genetic and molecular epidemiologic characteristics of 63 strains of Japanese encephalitis virus (JEV) isolated in Yunnan province, China during 1977-2010. Methods: Suckling mice were inoculated with viruses continuously and the viral nucleic acid were extracted from the brain-grinding supernatants of the infected and moribund mice, then the gene fragments of E region were amplified by RT-PCR. Bioinformatics (Clustal X, DNAstar, Mega 5.0 and other software) was used to analyze the nucleotide and deduced amino acid sequences and phylogenetic trees. Results: Yunnan strains of JEV could cause illness and deaths in suckling mice. The results of virus nucleic acid detection and sequencing indicated that nucleotide sequences of E gene of the 63 virus strains were obtained. Phylogenetic tree and homology analyses based on E genomes showed that 47 strains of the experimental virus belonged to genotype 1 (G-1) and 16 strains belonged to genotype 3 (G-3). The 47 isolates of G-1 were divided into 2 clades, of them, the earliest isolates of G-1 (M28, 1977 and BN82215, 1982) in Yunnan of China and the early isolates of G-1 (U70416, 1982; DQ084229, the year is unknown) in Thailand were in one clade, and the isolates of G-1 from 2007-2010 in Yunnan could be divided into 2 subgroups. The 16 isolates of G-3 from Yunnan were divided into 3 clades, among them, the isolates from 1970-1990s in Yunnan were in two clades, and the isolates from 2004 in Yunnan were in one clade. In addition, their main amino acid sites of antigenicity, pathogenic, virulence of both G-1 and G-3 had no significant change. Conclusion: JEV G-1 and G-3 co-circulated in Yunnan, and G-1 was predominant. The JEV strains isolated in different years and areas in Yunnan had different molecular epidemiologic characteristics and genetic diversity. The results of this study suggested that JEV G-1 might originate from Yunnan of China and adjacent Southeast Asia region.