丙型肝炎病毒体外感染MT—2细胞的研究

• 作 者: 陈智 刘勇 陈明华 沃健尔 刘克洲 何南祥 ＤＥＮＮＩＮＲｅｉｎｈａｒｄＨ ＲＥＩＮＨＡＲＤＵｒｓａ
• 机 构: 浙江大学医学院附属第一医院传染病研究所!３１０００３,浙江大学医学院附属第一医院传染病研究所!３１０００３,浙江大学医学院附属第一医院传染病研究所!３１０００３,浙江大学医学院附属第一医院传染病研究所
• 出 处: 中华肝脏病杂志
• 发表日期: 1999年03期
• 关键词: 丙型肝炎病毒 MT 体外感染 复制中间体 丙型肝炎患者 阴性血清 复制状态 持续阳性 特异性序列 扩增产物

目的建立体外丙型肝炎病毒感染的ＭＴ－２细胞模型。方法用ＨＣＶＲＮＡ阳性的丙型肝炎患者血清感染人淋巴细胞株ＭＴ－２以ＨＣＶＲＮＡ阴性血清作对照，用ＲＴ－ＰＣＲ分析ＨＣＶ在体外细胞培养中的复制状态。结果用ＨＣＶ阳性血清感染的ＭＴ－２细胞在培养第７天开始检出ＨＣＶＲＮＡ，并一直持续阳性至第２８天；培养至第１０天可测出ＨＣＶ复制中间体，于第１８天消失。用ＨＣＶ阴性血清感染的ＭＴ－２细胞未检出ＨＣＶＲＮＡ。扩增产物经Ｓｏｕｔｈｅｒｎ转移后用ＨＣＶ探针杂交证实ＨＣＶ特异性序列。结论ＨＣＶ可在ＭＴ—２中短期复制。

Objective To establish a MT-2 cell model infected by hepatitis C virus in vitro. Methods HCV RNA negative serum was used to infect human lymphocyte cell line MT-2 infected with HCV RNA positive hepatitis C virus, and the replication status of HCV in in vitro cell culture was analyzed by RT-PCR. Results HCV RNA was detected in HCV-positive sera of MT-2 cells on the 7th day of culture and continued to be positive on the 28th day. HCV replicative intermediates were detected on day 10 and disappeared on the 18th day. HCV RNA was not detected in MT-2 cells infected with HCV-negative sera. Amplification products were Southern blot followed by HCV probe hybridization to confirm HCV specific sequences. Conclusion HCV can be replicated in MT-2 in the short term.