【摘要】 目的 建立HBV阳性血清体外感染HepG2细胞的实验模式。 方法 在六孔板上培养HepG2细胞，用HBV阳性血清体外感染HepG2细胞，设立感染组和对照组。用HBV阳性血清和感染组细胞共孵育24h，0.01mol/L PBS清洗细胞后，每孔板中加入4ml DMEM细胞培养液，每隔12h收集细胞上清液至PBS洗后84h。收集完最后一次细胞上清液，收集细胞培养板中的细胞。ELSIA检测细胞培养上清中HBsAg;PCR方法检测细胞培养上清和细胞中HBV DNA;免疫荧光定量PCR检测细胞培养上清中HBV DNA的含量。 结果 HBV感染组细胞培养上清，在PBS洗后第12h，ELISA即可检测到HBsAg并持续到84h，PCR检测感染组细胞培养上清和感染组细胞的HBV DNA均阳性。荧光定量PCR检测感染组细胞培养上清中HBV DNA的结果:感染组PBS洗后(0h)的HBV定量为阴性，而在PBS洗后36h、84h细胞培养上清中HBV的量达到5×10 5 ，3×10 6 copies/ml)。 结论 用HBV阳性血清体外感染HepG2细胞的实验是可行的。

    【关键词】 乙型肝炎病毒;体外感染;细胞培养

    Study on hepatitis B virus infection of HepG2cell line in vitro

    WANG An-hui，XU De-zhong，MEN Ke，YAN Yong-ping，ZHANG Jing-xia，LU Juan.

    Department of Epidemiology，Fourth Military Medical University，Xi'an 710032，China

    【Abstract】 Objective To find out a suitable model of HBV positive serum infecting HepG2cell line.Methods HepG2cells were seeded on six-well cluster dishes.After24h plating，HepG2cells were cultured with HBV positive serum in infection group and with HBV negative serum in control group.24h later the inoculums were removed and the cells were extensively washed with0.01mol.L -1 phosphate-buffered saline(PBS).After PBS washed ......