Objective：This study aims to identify human herpesvirus 6（HHV-6） binding partner for glycoprotein 96（gp96），and analyze the effect of their interaction on HHV-6 infection. Methods：Immunoprecipitation，Western blot and confocal microscopyassays were used to identify and verify the interaction between gp96 and HHV-6 glycoprotein B（gB）. Effect of Gp96 on gB maturation was analyzed using exosome samples. Results：Immunoprecipitation results showed that HHV-6 gB interacted with gp96；gB and gp96 were colocalized，which was detected by confocal microscopy assays. Western blot results confirmed that overexpression of gp96 affected the maturation of gB. Conclusion：The gp96 is a binding partner for HHV-6 gB，and gp96 affects the maturation process of gB.