Construction of a pseudovirus reporting system for anti-West Nile virus drug screening / 国际药学研究杂志

ABSTRACT

Objective: To establish a pseudovirus reporting system was established for anti-WNV drug screening in order to by-pass security risk caused by live viruses in West Nile virus(WNV)research. Methods: 293T Cells were co-transfected with WNV replicon recombinant plasmid prWNV-Rluc containing Renilla luciferase reporter gene and recombinant plasmid pcDNA3.1-CME expressing WNV structural proteins C, M and E;and the cell culture supernatant containing pseudovirus was obtained at 72 h post-cotransfection. Results: After pseudovirus infection with the supernatant, BHK21 cells produced fluorescence with the intensity that increased dose-dependently with the supernatant containing pseudovirus. The WNV NS1 Protein expressed in the infected BHK21 cells was identified by Western blot, confirming the presence of WN pseudovirus in the supernatant. Neutralization assay showed that the WNV neutralizing antibody could effectively block the infection of BHK21 cells by the WN pseudovirus in the supernatant. Conclusion: The West Nile pseudovirus reporting system established in this study might be used without the biosafety level 3 laboratory (BSL-3) and could be applied to the screening of anti-WNV drugs.