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目的 :用丙型肝炎 (丙肝 )阳性血清 (抗HCV阳性及HCVRNART PCR阳性 )感染HepG2细胞 ,以期建立稳定的HCV感染的细胞模型。方法 :采用丙肝阳性血清连续感染法感染HepG2细胞 ,用逆转录多聚酶链反应检测传代HepG2细胞内HCVRNA正链和负链 ,电镜观察感染后的细胞内的病毒颗粒 ,免疫组化方法检测感染后的细胞内丙肝病毒蛋白抗原 (HCVNS5、HCVcapsid)的存在。结果 :感染后传代的第一代至第三代细胞内检出HCVRNA正链及负链 ;在感染后传代的细胞内发现球形、密集分布的类丙肝病毒颗粒 ,直径为 30～ 6 0nm ;免疫金银染色发现部分细胞内有聚集的银染颗粒 ;免疫电镜检查在细胞胞浆的囊泡样结构中发现聚集的金颗粒。结论 :HCV可在HepG2细胞内复制 ,可形成病毒颗粒 ,可随细胞传代。本研究初步建立了用丙肝阳性血清感染HepG2细胞的丙肝病毒感染细胞模型
OBJECTIVE: To infect HepG2 cells with positive hepatitis C (HCV) positive and positive HCVRN PCR PCR in order to establish a stable cell model of HCV infection. Methods: HepG2 cells were infected by continuous positive hepatitis C virus infection, positive and negative HCVRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR), and the infected cells were observed by electron microscopy. The expression of HCV RNA in infected HepG2 cells was detected by immunohistochemistry The presence of intracellular hepatitis C virus protein antigen (HCV NS5, HCVcapsid). Results: Positive and negative HCVRNA were detected in passage 1 to passage 3 cells after infection. Spheroid and densely distributed hepatitis C virus particles were found in the passaged cells after infection, with a diameter of 30-60 000 nm. Gold and silver staining found that some of the cells in the accumulation of silver-stained particles; immunoelectron microscopy in the cytoplasm of vesicular-like structure found in the accumulation of gold particles. Conclusion: HCV replicates in HepG2 cells and forms virus particles, which can be passaged with cells. This study initially established HepG2 infected Hepatitis C virus infected cell model with hepatitis C positive serum