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Article in Chinese | WPRIM | ID: wpr-329537


Objective To isolate and identify arboviruses from mosquito pools in some regions of Liaoning province.Methods Mosquitoes were collected from Shenyang,Yingkou,Panjin,Jinzhou and Dandong cities of Liaoning province in 2006.Viruses were isolated by inoculating the specimens onto C6/ 36 and BHK-21cells.The new isolates were identified using serological and molecular biological methods.Results 5410 mosquitoes were collected from the five cities in total.Three isolates produced CPE in C6/ 36 cell and five isolates produced CPE in both C6/36 and BI-IK-21 cell.Three isolates (LN0684,LN0688 and LN0689) were identified as Banna virus and one isolate (LN0636) was identified as Getah virus.Phylogenetic analysis showed that the three Banna virus strains were clustered into the same evolution branch as the other Chinese isolates.The identity of nucleotide sequence was between 91.2% and 94.7%,compared with other Banna virus strains.The new isolated Getah virus was clustered into the same branch with the strain of South Korea (swine).The identity of nucleotide sequence was 99.2%,when comparing with the strain of South Korea and was 95% to 99% with the strains fi'om Russia,mainland of China and Taiwan region.Conehmion Eight virus isolates,including three Banna virus,one Getah virus and four unknown virus strains were isolated from mosquitoes in Liaoning province.Banna virus and Getah virus were reported for the first time in Liaoning province,while Getah virus showed the highest nucleotide homology with the South Korea strains.

Chinese Journal of Virology ; (6): 173-177, 2009.
Article in Chinese | WPRIM | ID: wpr-334755


5 strains of virus isolated from Culex tritaeniorhynchus, Anopheles sinensis and Armigeres subalbatus, which caused cytopathic effect in C6/36 cells, had been obtained in the survey of arboviruses in Northwestern Yunnan Province. China. The virus particles displayed 70 nanometers diameter (n=7) with no envelope but spikes on the surfaces. RNA-PAGE of the genomes of the isolates showed 6-5-1 profile. A fragment of the 12th segment sequence was amplified by a pair of specific primers for Kadipiro virus strain JKT-7075 in RT-PCR. The full length of the 12th segment was 758 nucleotides, BLAST analysis revealed the highest identity was 90% to JKT-7075. Phylogenetic analysis demonstrated that the isolates appeared to be Kadipiro viruses (Family Reoviridae). It was the first report of kadipiro virus isolation in China.

Amino Acid Sequence , Animals , Anopheles , Virology , Cell Line , China , Coltivirus , Classification , Genetics , Culex , Virology , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
Article in Chinese | WPRIM | ID: wpr-325575


<p><b>OBJECTIVE</b>To sequence PrM and E gene of the Japanese encephalitis virus isolated from Gansu province in 2008 and analysis the genotype of new JEV isolates and the molecular characterization of E gene.</p><p><b>METHODS</b>Computer software was used to analyze nucleic acid sequence and deduced amino acid sequence, and draw phylogenetic trees, including ClustalX2.09, MegAlign and Mega4.</p><p><b>RESULTS</b>The six JEV strains were clustered in genotype I. 87.5%-87.9% identity in nucleotide sequence and 96.8%-97.2% identity in amino acid sequence were found in E gene when compared with the vaccine strain SA14-14-2. Eleven common amino acid differences were observed in E protein between new isolates and the vaccine strain.</p><p><b>CONCLUSION</b>Genotype I JEVs were isolated from mosquitoes collected in Gansu province. The amino acid difference occurred in sites that were not the key ones affecting the antigenic of JEV.</p>

Amino Acid Sequence , Animals , Cell Line , China , Cricetinae , Encephalitis Virus, Japanese , Chemistry , Classification , Genetics , Encephalitis, Japanese , Virology , Humans , Molecular Sequence Data , Phylogeny , Sequence Alignment , Viral Proteins , Chemistry , Genetics
Article in Chinese | WPRIM | ID: wpr-242645


<p><b>OBJECTIVE</b>To probe the primary characteristic of 0507JS11 virus isolated from Culex sp. and determine the classification of 0507JS11 virus in taxonomy.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>0507JS11 virus is a new member in Brevidensovirus.</p>

Animals , Culex , Virology , DNA, Viral , Genetics , Densovirinae , Classification , Genetics , Genome, Viral , Sequence Analysis, DNA
Article in Chinese | WPRIM | ID: wpr-254134


<p><b>OBJECTIVE</b>To isolate Japanese encephalitis virus (JEV) from mosquitoes collected in Liaoning province and analysis the genotype of new isolated JEV strains and the characters of nucleotide and amino acid in the E gene.</p><p><b>METHODS</b>Collected mosquitoes in Dandong Liaoning Province in August, 2006. Virus isolation was using issue culture cells. Isolated viruses were identified by using serological and molecular methods.</p><p><b>RESULTS</b>Two new JEV strains, LNDG07-02 and LNDG07-16, were isolated from 1500 mosquitoes were belonging to genotype 1. The identity of nucleotide and amino acid of E gene between new JEV strains and live attenuated vaccine strain SA14-14-2 were 87.8-88% and 97.2%, respectively. Total 11 amino acid sites were differences in E gene between new isolates and SA14-14-2. However, there were no differentiation between the new JEV strains and the isolates in Donggang 2002.</p><p><b>CONCLUSION</b>Genotype 1 JEV was isolated again from Donggang, since the first isolation of this genotype in 2002. Genotype 1 JEV continues in existence in Donggang Liaoning Province.</p>

Amino Acid Sequence , Animals , Brain , Pathology , Virology , Cell Line , China , Epidemiology , Culicidae , Virology , Encephalitis Virus, Japanese , Classification , Genetics , Encephalitis, Japanese , Epidemiology , Virology , Female , Genotype , Insect Vectors , Virology , Male , Mice , Molecular Sequence Data , Phylogeny , RNA, Viral , Genetics , Sequence Analysis, DNA
Chinese Journal of Virology ; (6): 270-275, 2007.
Article in Chinese | WPRIM | ID: wpr-334898


To compare the molecular characteristics of the Capsid gene and 3' untranslation region (3'UTR) of Getah viruses (GETV) isolated in Hainan Province and Hebei Province of China,the viral RNAs were extracted from M1(Hainan), HB0215-3 and HB0234(Hebei) virus stocks. Capsid gene segments and 3' UTR segments from three strains of Chinese GETV were obtained by RT-PCR and then sequenced. The obtained nucleotide sequences were analyzed using the Clustal X(1.8), DNASTAR, MAGA3.1 programs. The full-length Capsid gene of the 3 strains of Chinese GETV were comprised of 801, 804 and 804 nucleotides each, encoding the protein of 267,268 and 268 amino acids each. Sequencing of Capsid gene fragments showed that two strains of Hebei isolates were identical and had homology of 97.6% at nucleotide level and 97.8% at amino acids level with M1. Their homologies when compared with strains isolated from other countries were also high at nucleotide levels (95.4%-99.6%). The 3'UTR from the three strains were comprised of 411, 401 and 401 nucleotides each, and had found specific deletion of 10 nt at position 44-54 and two specific nucleotide sites that was T at position 64 and C at position 148. GETV isolated in China presented relation of the year of virus isolation with the phylogenesis distance when compared with the other GETV strains and comprised a genetically highly conserved group.

3' Untranslated Regions , Genetics , Alphavirus , Classification , Genetics , Base Sequence , Capsid Proteins , Genetics , China , Molecular Sequence Data , Phylogeny , Sequence Homology, Nucleic Acid