Objective Modeling HAdV-3 infect Hep-2 cells in vitro.The effect of realgar nanoparticles on the expression of HAdV-3 is detected by using fluorescent quantitative PCR.Method The experiment is divided into four groups:Hep-2 cells control group,HAdV-3 virus control group,realgar nanoparticle group and ribavirin group.In order to detect HAdV-3 viral load,add realgar nanoparticles and ribavirin in vitro and remain that vitro for 24 hours when HAdV-3 has infected Hep-2 cells,extract total DNA of Hep-2 cells infected by HAdV-3,and establish Real-time PCR reaction system of every experimental groups.Result The Hep-2 cells group has no amplification curve,the Ct value is greater than 35,which illustrate HAdV-3 pathogen detection is negative.However,realgar nanoparticles group,ribavirin group and the HAdV-3 group have amplification curve,the Ct values are 29.30 ± 0.08,33.05 ± 1.29,26.01 ± 0.25 respectively,which illustrate HAdV-3 pathogen detection is positive.The viral copy amount of the adenovirus group(66 699 932 ±23.85) is more than that of realgar nanoparticles group (912 435.44 ± 16.57),and much greater than that of ribavirin group(459 124.84 ± 12.82) (P ＜ 0.05).Conclusion The model of Hep-2 cell infected by HAdV-3 is reliable.The method of quantitative PCR is sensitive and specific.Realgar nanoparticles have a certain inhibition role for adenovirus nucleic acid replication.