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Objective To analyze the molecular characteristics of Japanese encephalitis virus (JEV) isolated in Northwestern Yunnan province, and to clarify the differences between the strains isolated in Northwestern and other parts of Yunnan province. Methods PrM, E and 3' untranslated region nueleotide acid sequences of the isolates were amplified by RT-PCR and then sequenced. Sequence alignment and phylogenetic analysis were performed by using Clustal 1.8X, DNASTAR, GENEDOC and Mega 3.1 programs. Results 12 of the 13 isolates of JEV obtained in Northwestern Yunnan were identified as genotype Ⅰ, only one strain was genotype Ⅲ of JEV. The 12 strains of genotype Ⅰ were clustered in different branches with other isolates obtained in other parts of Yunnan province. Data from sequence analysis on E gene found that the nucleotide identity was 0.2%-13.9% between the Northwestern isolates and other Yunnan strains. There were two kinds of nucleotides deletion patterns at 3' untranslated region with three and one deletions was found after termination eodon in genotype Ⅰ and Ⅲ isolates, respectively. Conclusion There were two genotypes of Ⅰ and Ⅲ in 13 strains of JEV in this study and genotype Ⅰ isolates were predominant (12/13). There were no apparent differences in E gene sequence between isolates obtained in the Northwestern and other parts of Yunnan. Three deletions were found in 3' untranslated region in genotypes Ⅰ isolates and one deletion was in genotypes Ⅲ.
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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.
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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.
Asunto(s)
Animales , Secuencia de Aminoácidos , Anopheles , Virología , Línea Celular , China , Coltivirus , Clasificación , Genética , Culex , Virología , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADNRESUMEN
<p><b>OBJECTIVE</b>To investigate arboviruses in some regions of Shanxi province, isolation and identification for arbovirus activity from mosquitoes was conducted.</p><p><b>METHODS</b>Mosquitoes were collected from these area in 2007 and then used for virus isolation by cell culture. The virus isolates were identified by molecular biology and the sequences were analyzed by bioinformatics.</p><p><b>RESULTS</b>Ten Banna virus (strains SX0765, SX0766, SX0767, SX0771, SX0789, SX0790, SX0793, SX0794, SX0795, SX0796) were isolated, and two Liaoning virus were also isolated from isolates SX0771, SX0794. Phylogenetic tree of the Banna virus isolates showed that ten strains are located in a distinct branch from all of the other Chinese Banna virus isolates. The homology is between 89.7% and 94.1%.</p><p><b>CONCLUSION</b>Ten Banna virus and two Liaoning virus were isolated during this arbovirus investigation in Shanxi province. New Banna virus isolates showed a distinct phylogenetic relationship with the other Chinese Banna virus strains.</p>
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Animales , Arbovirus , Clasificación , Genética , Línea Celular , China , Culicidae , Virología , Insectos Vectores , Virología , Datos de Secuencia Molecular , FilogeniaRESUMEN
<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>