Lack of genomic reassortment following infection of infant rats with group A and group B rotaviruses.
Lack of genomic reassortment following infection of infant rats with group A and group B rotaviruses.
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幼鼠感染 A 组和 B 组轮状病毒后缺乏基因组重排。
DOI：
10.1093/infdis/158.5.1120
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Vonderfecht,S
中科院分区：
文献类型：
--
作者：
Yolken,R;Arango-Jaramillo,S;Eiden,J;Vonderfecht,S
材料与方法制备B组轮状病毒IDIR株，按前文所述在幼鼠体内传代。A组轮状病毒MMU毒株（血清型3）最初从恒河猴中分离出来，由Richard Wyatt博士（马里兰州贝塞斯达国立卫生研究院）提供。使用先前描述的方法[2]在MA-104细胞（MA Bioproducts, Walkersville, Md）中繁殖病毒。采用先前描述的方法制备MMU病毒株的中和单克隆抗体和豚鼠血清超免疫[2,3]。3 - 5日龄大鼠（Charles River Laboratories, Portage, michigan）口服106 pfu MMU和标准接种B组轮状病毒idr的混合物（第一次传代）；观察他们是否发展成腹泻病。分别于感染后第1、2、4天采集2 ~ 4只感染大鼠肠道内容物。选择这些天是因为在感染A组轮状病毒[12]的动物中观察到体内重组的时间过程。在相同的条件下，将一部分肠道内容物汇集并喂给另一组幼鼠（第二段）。这一过程也导致了腹泻病的产生。在1 ng /mL胰蛋白酶的作用下，检测两组大鼠肠道内容物在MA-104细胞上斑块的生成。在0.5 Vo琼脂糖覆盖层中添加三种不同的MMU中和单克隆抗体，在存在或不存在的情况下培养肠道内容物。接种后第三天添加含有0.5^ 0琼脂糖和中性红色的第二次覆盖层，斑块被视为不占用中性红色染色的离散区域。
Materials and MethodsThe IDIR strain of group B rotavirus was prepared and passaged in infant rats as previously described [9]. The MMU strain of group A rotavirus (serotype 3) was origi-nally isolated from rhesus monkeys and was obtained from Dr. Richard Wyatt (National Institutes of Health, Be-thesda, Md). The virus was propagated in MA-104 cells (MA Bioproducts, Walkersville, Md) by using previously described methods [2]. Neutralizing monclonal antibodies and guinea pig sera hyperimmune to the MMU strain of virus were prepared by means of previously described methods [2, 3]. Three-to five-day-old rats (Charles River Laboratories, Portage, Mich) were orally inoculated with a mixture of 106 pfu of MMU and a standard inoculum of the group B rotavirus IDIR (first passage); they were observed for the development of diarrheal disease. Intes-tinal contents of two to four infected rats were collected on days 1, 2, and 4 after infection. These days were selected because of the time course of in vivo reassortment observed in animals infected with group A rotaviruses [12]. A portion of the intestinal contents was pooled and fed to an-other group of infant rats under identical conditions (second passage). This procedure also resulted in the generation of diarrheal disease. The intestinal contents of both groups of rats were tested to determine their generation of plaques on MA-104 cells in the presence of 1 ng of trypsin/mL. The intestinal contents were cultured in the presence or absence of three different neutralizing monoclonal antibodies to MMU added to an 0.5 Vo agarose overlay. A second overlay containing 0.5^ 0 agarose and neutral red was added three days after inoculation, and plaques were visualized as discrete areas that did not take up the neu-tral red stain.