ISOLATION AND SEROTYPIC CHARACTERIZATION OF HUMAN AND ANIMAL ROTAVIRUSES

轮状病毒现在可以通过技术在细胞培养中很容易地生长 最初由日本调查人员描述，包括 胰酶对病毒的前处理，将胰酶掺入 维护介质以及在辊上培养辊管的方法 仪器。我们在这个项目中的目标有三个：(1) 在细胞培养中直接培养各种人和动物 来自不同地理区域的轮状病毒株和 (2)定义不同群体间的血清型多样性和相似性 这些病毒基于其VP4和VP7的特异性；以及 选择和开发潜在的轮状病毒候选疫苗(包括 直接细胞培养分离物或实验室生产的重配子 菌株)。在本项目中，轮状病毒分离株的血清型为 以空斑减少中和试验(PRN)为特征。 三个单一的VP7基因替换重组子是用 猪轮状病毒戈特弗里德株(VP7 4型)和人轮状病毒 毒株D(VP7血清型1)、DS-1(VP7血清型2)或M(VP7血清型 3)，每一株仅具有D、DS-1或M的VP7基因 剩下的10个基因来自戈特弗里德菌株。 针对重组菌株制备的高免豚鼠抗血清 戈特弗里德x DS-1，不仅中和了亲本菌株戈特弗里德 和DS-1，也包括属于VP7 1型(WA)的人轮状病毒 株)、3株(P株)或4株(VA70和ST3株)。

Rotavirus can now br grown readily in cell cultures by techniques originally described by Japanese investigators which include pre-treatment of virus with trypsin, incorporation of trypsin in the maintenance medium, and incubation of roller tube cultures on a roller apparatus. Our objectives in this project are three-fold: (1) to cultivate directly in cell cultures a variety of human and animal rotavirus strains derived from diverse geographical areas and populations; (2) to define serotypic diversity and similarity among these viruses based on their VP4 and VP7 specificities; and (3) to select and develop potential rotavirus vaccine candidates (including direct cell-culture isolates or laboratory produced reassortant strains). In this project, the serotype of rotavirus isolates was characterized by plaque-reduction neutralization (PRN) assay. Three single VP7 gene substitution reassortants were generated using a porcine rotavirus Gottfried strain (VP7 serotype 4) and human rotavirus strains D (VP7 serotype 1), DS-1 (VP7 serotype 2), or M (VP7 serotype 3), each of which possesses only the VP7 gene of D, DS-1, or M with the remaining ten genes being derived from the Gottfried strain. Hyperimmune guinea pig antiserum raised against the reassortant strain Gottfried x DS-1, neutralized not only the parental strains Gottfried and DS-1 but also human rotaviruses belonging to VP7 serotype 1 (Wa strain), 3 (P strain), or 4 (VA70 and ST3 strains).