Kanto T, Hayashi N, Takehara T, Hagiwara H, Mita E, Naito M, Kasahara A, Fusamoto H, Kamada T
First Department of Medicine, Osaka University Medical School, Japan.
Hepatology. 1994 Feb;19(2):296-302.
Hepatitis C virus is reported to have a low buoyant density in sucrose. To determine the density of hepatitis C virus in the circulation of infected hosts and its association with the degree of liver inflammation, we examined serum samples from 10 patients who were positive for both hepatitis C virus antibody (C100 antigen) antibody and serum hepatitis C virus RNA. After the serum was ultracentrifuged in sucrose density gradient (10% to 60%), the hepatitis C virus RNA titer in each collected fraction was quantified by means of competitive reverse transcription-polymerase chain reaction. In samples from five blood donors, the hepatitis C virus RNA titer had a single peak at fractions with densities of 1.08 to 1.11 gm/ml. In samples from five patients with ALT abnormalities, the titer had two peaks at fractions with 1.09 to 1.10 gm/ml and 1.22 to 1.25 gm/ml. After the selected samples were treated with detergents and ultracentrifuged, the titer in the 1.08 to 1.11 gm/ml fractions decreased and that in the 1.22 to 1.25 gm fractions increased. This result implied that the hepatitis C virus density changed with removal of the viral envelope by lipid solvents. Thus the buoyant density of hepatitis C virus in sucrose was 1.08 to 1.11 gm/ml for an intact virion and 1.22 to 1.25 gm/ml for what was presumed to be a nucleocapsid. These results demonstrated that HCV virion is a dominant form in the circulation of blood donors without ALT abnormalities. In patients with liver inflammation HCV particles with higher densities of 1.22 to 1.25 gm/ml coexist with virion in the circulation, which might be presumed nucleocapsids.
据报道,丙型肝炎病毒在蔗糖中的浮力密度较低。为了确定丙型肝炎病毒在受感染宿主循环系统中的密度及其与肝脏炎症程度的关系,我们检测了10例丙型肝炎病毒抗体(C100抗原)和血清丙型肝炎病毒RNA均呈阳性的患者的血清样本。血清在蔗糖密度梯度(10%至60%)中进行超速离心后,通过竞争性逆转录-聚合酶链反应对每个收集组分中的丙型肝炎病毒RNA滴度进行定量。在5名献血者的样本中,丙型肝炎病毒RNA滴度在密度为1.08至1.11克/毫升的组分处有一个单峰。在5名谷丙转氨酶(ALT)异常患者的样本中,滴度在密度为1.09至1.10克/毫升和1.22至1.25克/毫升的组分处有两个峰。对选定样本用去污剂处理并超速离心后,1.08至1.11克/毫升组分中的滴度下降,而1.22至1.25克/毫升组分中的滴度上升。这一结果表明,丙型肝炎病毒密度随着脂质溶剂去除病毒包膜而发生变化。因此,完整病毒体的丙型肝炎病毒在蔗糖中的浮力密度为1.08至1.11克/毫升,而推测为核衣壳的丙型肝炎病毒在蔗糖中的浮力密度为1.22至1.25克/毫升。这些结果表明,在没有ALT异常的献血者循环系统中,HCV病毒体是主要形式。在有肝脏炎症的患者中,密度为1.22至1.25克/毫升的较高密度HCV颗粒与病毒体在循环系统中共存,这些颗粒可能是推测的核衣壳。