鲤疱疹病毒Ⅱ型的理化及生物学特性和超微形态发生

为了查明鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2,Cy HV-2)的理化与生物学特性以及病毒在细胞内的超微形态发生过程,利用新建立的对Cy HV-2敏感的异育银鲫脑组织细胞系(Gi CB),对Cy HV-2的理化及生物学特性进行了详细研究,比较了不同来源鱼类细胞系对Cy HV-2感染的敏感性,并对体外培养细胞中Cy HV-2病毒粒子及其超微形态发生过程进行了电镜观察。结果显示,Cy HV-2对热、酸、碱、有机溶剂和冻融敏感;常用鱼类细胞系EPC、RTG-2、Koi-Fin、CIK、CCK、PF-Fin对Cy HV-2的感染不敏感,特异性巢式PCR检测盲传至第7代Cy HV-2细胞培养物,结果均为阴性;Cy HV-2在Gi CB细胞中的增殖动态研究结果表明:病毒感染细胞经过12 h的隐晦期,24 h开始进入对数生长期,96 h病毒滴度达到最高值(107.52±0.26 TCID50/m L),然后进入平台期;透射电子显微镜观察结果显示,Cy HV-2感染细胞可分为吸附与侵入、复制与装配、成熟与释放3个主要过程,病毒进入对数生长期后,被感染细胞内可见形态典型的疱疹病毒颗粒。

The physical-chemical and biological characteristics of Cyprinid herpesvirus 2 and its ultrastructural morphogenesis in vitro

The aim of the present study was to investigate the physical-chemical, biological characteristics Cyprinid herpesvirus 2 (CyHV-2) and its ultrastructural morphogenesis in vitro. By using the newly established cell line derived from gibel carp brain, designated GiCB, which was highly permissive to the infection of CyHV-2, the physical-chemical and biological characteristics, growth curve of CyHV-2 were investigated; meanwhile, the susceptibility of available fish cell lines to CyHV-2 infection was compared and the ultrastructural morphogenesis was conducted in this study. The results indicated that the CyHV-2 is sensitive to the treatment of heat, acid (pH 3.0), alkali (pH10.0), chloroform, ether and frozen-thawed. CyHV-2 could not propagated in fish cell lines EPC,RTG-2,Koi-Fin,CIK,CCK andPF-Fin, etc., which were verified by CPE observation and CyHV-2 DNA Nest-PCR assay; According to the growth curve of CyHV-2 in GiCB cells, a log phase increase in virus production was followed by an eclipse period of about 12h. Virus production peaked around 96h (107.52±0.26 TCID50/ml). The CyHV-2 ultrastructural morphogenesis in GiCB cells presented in three main stages, including virus adsorption and entry, replication and assembly, maturation and budding. This study provided useful information for the further studies on CyHV-2 infection mechanisms, pathogenicity and the control methods of the disease caused by the virus.