Rodriguez L L, Owens J H, Peters C J, Nichol S T
Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
Virology. 1998 Mar 1;242(1):99-106. doi: 10.1006/viro.1997.8990.
In order to determine the frequency and characteristics of reassortment among viruses causing hantavirus pulmonary syndrome (HPS), mixed infections were initiated in tissue culture by using two closely related strains of Sin Nombre virus, CC107 (from eastern California) and NMR11 (from New Mexico), which share the same species of rodent host in nature, the deer mouse (Peromyscus maniculatus). Potential reassortant virus plaques were screened by multiplex RT-PCR, using primers specific for individual genome segments of each strain. Reassortant viruses involving the M and S segments and, to a lesser extent, the L segment were detected in 8.5% of 294 progeny plaques tested. In addition, approximately 30% of the progeny virus plaques appeared to contain S or M segments originating from both parental virus strains, i.e., they were diploid. Most of these diploid virus genotypes were not stable, becoming either reassortant or parental virus strains upon plaque-to-plaque virus passage. In contrast to the results above, only one virus reassortant and four diploids were observed among 163 progeny virus plaques from mixed infections between Sin Nombre virus NMR11 and the genetically more distant Black Creek Canal virus, an HPS-causing virus from Florida, which has the cotton rat (Sigmodon hispidus) as its natural host.
为了确定引起汉坦病毒肺综合征(HPS)的病毒之间重配的频率和特征,利用两种密切相关的辛诺柏病毒株CC107(来自加利福尼亚州东部)和NMR11(来自新墨西哥州)在组织培养中引发混合感染,这两种病毒在自然界中共享相同的啮齿动物宿主物种——鹿鼠(白足鼠)。通过多重逆转录聚合酶链反应(multiplex RT-PCR)筛选潜在的重配病毒噬斑,使用针对每个病毒株单个基因组片段的特异性引物。在检测的294个后代噬斑中,8.5%的噬斑中检测到涉及M和S片段以及程度较轻的L片段的重配病毒。此外,大约30%的后代病毒噬斑似乎含有来自两个亲本病毒株的S或M片段,即它们是二倍体。这些二倍体病毒基因型大多不稳定,在噬斑到噬斑的病毒传代后会变成重配病毒株或亲本病毒株。与上述结果相反,在辛诺柏病毒NMR11与遗传距离更远的黑溪运河病毒(一种来自佛罗里达州的引起HPS的病毒,其自然宿主是棉鼠)的混合感染产生的163个后代病毒噬斑中,仅观察到1个病毒重配体和4个二倍体。