ABSTRACT
To investigate the inhibitory effect of RNA interference (RNAi) on dengue virus I (DENV-1) replication. Small interfering RNA (siRNA) against the PreM gene of dengue virus was synthesized and transfected into C6/36 cells with liposome, which was then attacked by DENV-1 virus. The antiviral effect of siRNA was evaluated by cytopathic effect (CPE), the cell survival rate measured by MTT, and virus RNA quantified by real-time RT-PCR. The results showed that after 7 days post infection of dengue virus, the transfected C6/36 cells showed less CPE. The cell survival rate of the transfected C6/36 cells increased by 2.26 fold, and the amount of virus RNA in the transfected cells was reduced by about 97.54% as well. These findings indicated that the siRNA could effectively inhibit dengue virus RNA replication, and protect C6/36 cells from viral attack, indicating its potential role in prevention and treatment of dengue fever.
Subject(s)
Animals , Cell Line , Dengue Virus , Genetics , Physiology , RNA Interference , Physiology , RNA, Small Interfering , Genetics , RNA, Viral , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Virus Replication , Genetics , PhysiologyABSTRACT
To understand the genetic characteristics of hemagglutinin (HA) and neuraminidase (NA) of type B influenza viruses in Guangzhou in 2006, three virus strains from etiology surveillance and seven strains from outbreaks were investigated. Genome RNAs of type B influenza viruses were extracted and reverse-transcripted into cDNAs using random primers. The whole-length DNA of HA and NA were amplified by polymerase chain reaction (PCR), cloned into T-A plasmid and sequenced, and analyzed phylogenetically by DNAstar software. The results showed that the HA of type B influenza viruses were similar and the homology were more than 99%. The type B influenza viruses belong to Victoria lineage. The NA of the type B influenza viruses were similar and the homology were more than 98%. Phylogenetic trees of HA and NA showed that the isolates from etiology surveillance formed a cluster, and the isolates from outbreaks were separated from the cluster. The homology of the type B influenza viruses with B/Shanghai/361/2002, which is the WHO recommended influenza vaccine strain in 2005-2006, were 88.9%-89.7%. It suggested the protective effect of influenza vaccine against type B influenza viruses used in 2005-2006 in Guangzhou may not be afforded.