ISOLATION AND SEROTYPIC CHARACTERIZATION OF HUMAN AND ANIMAL ROTAVIRUSES

人和动物轮状病毒的分离和血清型特征

• 批准号: 3803153
• 负责人: Y HOSHINO
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3803153
• 关键词: Rotavirus; antibody neutralization test; cross immunity; fusion gene; genetic strain; growth media; human tissue; immunological substance; microorganism culture; rotavirus vaccines; serotyping; virion; virus classification; virus genetics; virus protein

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).

轮状病毒现在可以很容易地在细胞培养中生长， 最初是由日本研究人员描述的， 用胰蛋白酶预处理病毒，将胰蛋白酶掺入病毒中， 维持培养基，并在滚筒上孵育滚筒管培养物 设备. 我们在这个项目中有三个目标：（1） 直接在细胞培养物中培养各种人类和动物 来自不同地理区域的轮状病毒株， （2）确定血清型多样性和相似性 这些病毒基于它们的VP 4和VP 7特异性;和（3） 选择和开发潜在的轮状病毒候选疫苗（包括 直接细胞培养分离株或实验室生产的抑制剂 菌株）。 在本项目中，轮状病毒分离株的血清型为 通过空斑减少中和（PRN）测定表征。 使用一种抗VP 7基因的抗体产生三种单一VP 7基因取代的抑制剂。 猪轮状病毒Gottfried株（VP 7血清型4）和人轮状病毒 菌株D（VP 7血清型1）、DS-1（VP 7血清型2）或M（VP 7血清型 3），其中每一种仅具有D、DS-1或M的VP 7基因， 其余10个基因来自戈特弗里德菌株。 抗抗性菌株的超免疫豚鼠抗血清 Gottfried x DS-1，不仅中和了亲本菌株Gottfried 和DS-1，但也包括属于VP 7血清型1的人轮状病毒（Wa 菌株）、3（P菌株）或4（VA 70和ST 3菌株）。