Fritzemeier J, Greiser-Wilke I, Haas L, Pituco E, Moennig V, Liess B
Institute of Virology, Veterinary School Hannover, Germany.
Vet Microbiol. 1995 Sep;46(1-3):285-94. doi: 10.1016/0378-1135(95)00093-p.
Antigenic and genetic analyses were performed in order to establish relationships between the noncytopathogenic (ncp) and the cytopathogenic (cp) bovine viral diarrhoea viruses (BVDV) involved in the induction of a case of experimentally induced "late-onset" mucosal disease (MD) symptoms. The persistent ncpBVDV, the cpBVDV used for superinfection (strain TGAC) and the virus isolates from faeces (cpX) were examined using an immunoplaque test (IPT) to distinguish between cp and ncp virus populations. The cp populations were cloned by plaque purification and found to be free of ncpBVDV when using the IPT. The cpBVDV clones and the persistent ncpBVDV were analysed in an enzyme immunoassay on heat-fixed infected cells (IM-EIA) and in a neutralization test using a panel of 27 monoclonal antibodies against the E0 (gp48) and E2 (gp53) viral glycoproteins. It was found that strain TGAC contained two antigenically distinct subpopulations of cpBVDV (TGAC-B1 and TGAC-B2). The endogenous ncpBVDV and the cpX clones had the same reactivity pattern in both tests. In addition, p80 gene duplications in the genomes of the cpBVDV clones were analysed using the polymerase chain reaction and subsequent restriction enzyme analysis of the amplicons. The clones analysed from TGAC-B1 and those from cpX had gene duplications of identical sizes showing the same restriction enzyme patterns. Our results suggest that the cpBVDV which finally lead to "late-onset" MD arose by recombination and/or by mutations of the cpBVDV used for superinfection.
为了确定参与诱导一例实验性诱导的“迟发性”黏膜病(MD)症状的非致细胞病变(ncp)和致细胞病变(cp)牛病毒性腹泻病毒(BVDV)之间的关系,进行了抗原和基因分析。使用免疫斑试验(IPT)检测持续性ncpBVDV、用于重叠感染的cpBVDV(TGAC株)以及粪便中的病毒分离株(cpX),以区分cp和ncp病毒群体。通过噬斑纯化对cp群体进行克隆,使用IPT发现其不含ncpBVDV。在热固定感染细胞的酶免疫测定(IM-EIA)中以及使用一组针对病毒糖蛋白E0(gp48)和E2(gp53)的27种单克隆抗体进行的中和试验中,对cpBVDV克隆和持续性ncpBVDV进行了分析。发现TGAC株包含两个抗原性不同的cpBVDV亚群(TGAC-B1和TGAC-B2)。内源性ncpBVDV和cpX克隆在两项试验中具有相同的反应模式。此外,使用聚合酶链反应以及对扩增子进行后续限制性酶切分析,对cpBVDV克隆基因组中的p80基因重复进行了分析。从TGAC-B1分析的克隆和从cpX分析的克隆具有相同大小的基因重复,显示出相同的限制性酶切模式。我们的结果表明,最终导致“迟发性”MD的cpBVDV是通过用于重叠感染的cpBVDV的重组和/或突变产生的。