论文部分内容阅读
丁型肝炎(HDV)的病毒学研究表明HDV的感染必须在HBV的共感染情况下进行,故要研究HDV的体外感染必须制备一种理想的体外感染的组织培养系统。作者为了确定黑猩猩肝细胞原代培养能否作为体外感染HDV的靶细胞,采用了感染或未感染HBV黑猩猩的肝细胞,在无牛血清培养液中培养3天后,细胞接种含HDV与HBV的黑猩猩血清。此血清系用HDV克隆的cDNA感染已感染HBV的黑猩猩血清,并已测定出每毫升感染细胞分别含10~(?)与10~6的HDV与HBV颗粒。将每毫升含10~8HBV基因的血清作对照。接种3天后,每3天收获一次,检查细胞感染情况:抽提两种病毒的核酸并将其转染到HDV与HBV杂交探针的醋酸纤维素薄膜上,以测定感染肝细胞中HDV RNA与HBV DNA的复制情况。并用免
Virological studies of hepatitis D virus (HDV) have shown that HDV infection must be carried out in co-infections with HBV. Therefore, to study the in vitro infection of HDV, an ideal tissue culture system for in vitro infection must be prepared. To determine whether chimpanzee hepatocytes primary cultures can be used as target cells for in vitro infection of HDV, the authors used hepatocytes infected or not infected with HBV chimpanzees and cultured for 3 days in a bovine serum-free medium before inoculation of chimpanzees with HDV and HBV serum. This serum infects chimpanzee serum that has been infected with HBV with the cDNA of the HDV clone and has determined that the HDV and HBV particles, which have 10 ~ (?) And 10 ~ 6, respectively, per ml infected cells. Each ml of serum containing 10 ~ 8HBV gene as a control. Three days after inoculation, harvested once every three days and checked for cell infection: Two viral nucleic acids were extracted and transfected into HDV and HBV hybridization probe cellulose acetate films to determine the presence of HDV RNA in hepatocytes HBV DNA replication. And use free