ABSTRACT
Japanese encephalitis virus (JEV), a mosquito-borne zoonotic pathogen, is one of the major causes of viral encephalitis worldwide. Previous phylogenetic studies based on the envelope protein indicated that there are four genotypes, and surveillance data suggest that genotype I is gradually replacing genotype III as the dominant strain. Here we report an evolutionary analysis based on 98 full-length genome sequences of JEV, including 67 new samples isolated from humans, pigs, mosquitoes, midges. and bats in affected areas. To investigate the relationships between the genotypes and the significance of genotype I in recent epidemics, we estimated evolutionary rates, ages of common ancestors, and population demographics. Our results indicate that the genotypes diverged in the order IV, III, II, and I and that the genetic diversity of genotype III has decreased rapidly while that of genotype I has increased gradually, consistent with its emergence as the dominant genotype.
Subject(s)
Encephalitis Virus, Japanese/classification , Encephalitis Virus, Japanese/genetics , Encephalitis, Japanese/epidemiology , Encephalitis, Japanese/virology , Genome, Viral , Animals , Asia/epidemiology , Cluster Analysis , Encephalitis Virus, Japanese/isolation & purification , Genotype , Humans , Molecular Sequence Data , Phylogeny , RNA, Viral/genetics , Sequence Analysis, DNAABSTRACT
In 2006, the first Chinese Tahyna virus isolate (XJ0625) was obtained in Xinjiang province and human infection were found in the same region. In this study, cell culture, animal experiments, electron microscopy, immunofluorescence assay and cross neutralization tests were performed to see the cell susceptibility, animal pathogenicity, morphology and antigenic and other biological characteristics of XJ0625. In addition, molecular biology software was used to analyze the characteristics of molecular evolution. The results showed that BHK-21 cell line was susceptible to XJ0625 and the virus was lethal to suckling mice when injected by intracranial ways. Similar to the other Bunyavirus, Tahyna virus is spherical enveloped virus under electron microscopy. XJ0625 infected cells showed strong fluorescent signal and could be neutralized by immune asities fluid with immnity to protype Tahyna virus Bardos 92. The sequence of the S and M segments showed 91.8% and 81.9% homology with Bardos 92.
Subject(s)
Encephalitis Virus, California/genetics , Encephalitis Virus, California/isolation & purification , Phylogeny , Animals , Cell Line , China , Encephalitis Virus, California/physiology , Evolution, Molecular , Female , Fluorescent Antibody Technique , Mice , Nervous System/virology , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
OBJECTIVE: To probe the primary characteristic of 0507JS11 virus isolated from Culex sp. and determine the classification of 0507JS11 virus in taxonomy. METHODS: 0507JS11 virus was cultured in Aedes albopictus C6/36 cells and cytopathic effects (CPEs) were recorded. Electro-microscopic morphology of 0507JS11 virus was observed. Total DNA extract of 0507JS11 virus was detected by 1% Agarose Gel Electrophoresis. Complete genomic sequence of 0507JS11 virus was sequenced and then made phylogenetic analysis. RESULTS: 0507JS11 virus could cause CPEs in Aedes albopictus C6/36 cells. Viral particles have no envelope and appear icosahedron symmetry with diameter of 20 nm. The genome of 0507JS11 virus was positive single strand DNA (ssDNA) with full length of 3977 nt. However, a DNA band about 4 kbp was observed in the electrophoresis of total DNA extract of 0507JS11 virus. The coding region of the genome included three ORFs, ORF1 and ORF2 code NSP1 and NSP2, ORF3 codes VP. Phylogenetic analysis of the complete genomic sequence of 0507JS11 virus indicated an independent linear in Brevidensovirus. CONCLUSION: 0507JS11 virus is a new member in Brevidensovirus.
Subject(s)
Culex/virology , Densovirinae/classification , Densovirinae/isolation & purification , Animals , DNA, Viral/genetics , Densovirinae/genetics , Genome, Viral , Sequence Analysis, DNAABSTRACT
OBJECTIVE: To isolate viruses from mosquitoes in the south of Xinjiang and identify these viruses primarily. METHODS: A total of 13 491 mosquitoes were collected in the south of Xinjiang from Jul to Aug, 2005. These mosquitoes were divided into 130 groups and grinded respectively. The supernates were inoculated in C6/36 and Vero cells. Viruses isolated were detected, the genomic nucleic types by electrophoresis of viral genomes and the morphologies observed under electronmicroscope. RESULTS: All 42 viruses were isolated, which caused CPEs on C6/36 but not on Vero cells. 27 viruses showed similar genomic profiles with 12 dsRNA segments. 1 virus displayed genomic profile with 10 dsRNA segments. 5 viruses took on similar genomic profiles with about 4 kbp DNA band. 9 viruses did not get any taxonomy information. Electromicroscopic pictures of these viruses revealed that above four types of viruses had distinguished morphologies indicating different virus species. CONCLUSION: There should be several virus species in the mosquitoes in the south of Xinjiang. dsRNA virus with 12 genomic segments should play analysis a predominant role in the south of Xinjiang.
Subject(s)
Culicidae/virology , Genome, Viral , Insect Viruses/isolation & purification , Animals , Bluetongue virus/classification , Bluetongue virus/genetics , Bluetongue virus/isolation & purification , China , Chlorocebus aethiops , Dengue Virus/classification , Dengue Virus/genetics , Dengue Virus/isolation & purification , Insect Viruses/classification , Insect Viruses/genetics , RNA, Double-Stranded/genetics , RNA, Viral/genetics , Reassortant Viruses/genetics , Sequence Analysis, DNA , Vero CellsABSTRACT
In 2006, Tahyna virus was isolated from Culex spp. mosquitoes collected in Xinjiang, People's Republic of China. In 2007, to determine whether this virus was infecting humans, we tested serum from febrile patients. We found immunoglobulin (Ig) M and IgG against the virus, which suggests human infection in this region.
