Construction and identification of infectious model virus of AIV strain H5N1

[Objective] HA and NA of Avian influenza virus strain H5N1 were used to construct a model virus of AIV.The activity and specificity of the model virus were tested respectively.[Method] HA and NA genes of AIV H5N1 were amplified and subcloned into pcDNA4.0.The recombinant plasmids were named pcDNA-HA and pcDNA-NA respectively.Using calcium phosphate cotransfection method,these plasmids together with HIV backbone pNL4-3.Luc.R-E-were transfected into 293T cell lines.The model virus was named HIV/HA-NA.The activity and specificity of the model virus HIV/HA-NA were tested subsequently.[Result] Model viruses HIV/HA-NA were obtained after pcDNA-HA,pcDNA-NA and pNL4-3.Luc.R-E-was cotransfected into 293T cell lines.Specificity test with the sensitive cells and the activity test showed that HIV/HA-NA can mimic the infectious process of AIV.By the help of reporting system,the infectious activity of the model virus can be detected quantificationally.[Conclusion] The model virus HIV/HA-NA can mimic the infectious process of AIV and report the infectious extent quantificationally.The model virus is safe and reliable since it can not be proliferated.It can be used as an excellent tool to screen antibody and special medicine of AIV in common laboratory.