Sauerbrei A, Schacke M, Schultz U, Egerer R, Merkle I, Glebe D, Gerlich W, Wutzler P
Institute of Virology and Antiviral Therapy, Friedrich-Schiller University, Jena, Germany.
J Virol Methods. 2005 Nov;129(2):178-85. doi: 10.1016/j.jviromet.2005.05.025.
Hepatitis B virus (HBV) is an important virus used in disinfection procedures for blood spillage. However, validation of HBV inactivation is difficult, since there are no feasible infectivity assays. In some countries, the duck HBV (DHBV) is recognized as a suitable model for testing antiviral activity of chemical biocides against HBV. Currently, DHBV-infected ducks are required for preparation of the test virus as well as eggs from DHBV-free flocks for testing DHBV infectivity. To improve the practicality of the system, we suggested to use commercially available embryonated duck eggs for preparation of DHBV-susceptible hepatocyte cultures and to exclude infected hepatocytes by pre-screening with qualitative detection of DHBV DNA using polymerase chain reaction (PCR). A standardized DHBV test virus was prepared from the DHBV DNA-transfected hepatoma cell line D2, which contained 10(11)DHBV DNA molecules per mL detected by light cycler real-time PCR. Infection of cell cultures was most efficient 4 days after plating. The best identification of infected cultures was possible 6 days after infection with immunofluorescence using an antiserum against DHBV surface antigen.
乙型肝炎病毒（HBV）是用于血液溢出物消毒程序的一种重要病毒。然而，由于没有可行的感染性检测方法，HBV灭活的验证很困难。在一些国家，鸭乙型肝炎病毒（DHBV）被认为是测试化学杀菌剂对HBV抗病毒活性的合适模型。目前，制备测试病毒需要感染DHBV的鸭子，以及来自无DHBV鸡群的用于测试DHBV感染性的鸡蛋。为提高该系统的实用性，我们建议使用市售的鸭胚蛋制备对DHBV敏感的肝细胞培养物，并通过使用聚合酶链反应（PCR）定性检测DHBV DNA进行预筛选来排除受感染的肝细胞。从DHBV DNA转染的肝癌细胞系D2制备标准化的DHBV测试病毒，通过荧光定量PCR检测，每毫升含有10¹¹个DHBV DNA分子。接种后4天，细胞培养物的感染效率最高。感染后6天，使用抗DHBV表面抗原的抗血清通过免疫荧光法能够最好地鉴定受感染的培养物。
