Expression of Recombinant Capsid Proteins of Chitta Virus, a Genogroup II Norwalk Virus, and Development of an ELISA to Detect the Viral Antigen
Expression of Recombinant Capsid Proteins of Chitta Virus, a Genogroup II Norwalk Virus, and Development of an ELISA to Detect the Viral Antigen
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Chitta 病毒（一种基因组 II 诺瓦克病毒）重组衣壳蛋白的表达以及检测病毒抗原的 ELISA 的开发
DOI：
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发表时间：
2000
影响因子：
2.6
通讯作者：
N. Takeda
中科院分区：
文献类型：
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作者：
Shinichi Kobayashi;K. Sakae;Yasumoto Suzuki;H. Ishiko;K. Kamata;Kenji Suzuki;K. Natori;T. Miyamura;N. Takeda
克隆Chitta病毒（CHV）第二个开放阅读框（ORF2）基因，构建重组杆状病毒。 CHV ORF2 预计编码 535 个氨基酸 (aa) 的衣壳蛋白。 CHV 在衣壳区域与基因组 II 诺瓦克病毒 (NV) (65-85%) 显示出较高的氨基酸一致性，但与基因组 I NV (44-46%) 的氨基酸一致性较低。 ORF2 基因的系统发育分析表明，CHV 在遗传上与基因组 II NV 中的夏威夷病毒密切相关。当重组杆状病毒在昆虫细胞中表达时，CHV 重组衣壳蛋白 (rCHV) 自组装形成空病毒样颗粒 (VLP)。开发了一种基于 rCHV 抗血清的酶联免疫吸附测定 (ELISA)，用于检测粪便中的 CHV 抗原。抗原 ELISA 似乎对 rCHV 和 CHV 样毒株具有高度特异性。此外，结合使用抗原 ELISA，使用针对两种抗原不同的重组 VLP（重组千叶病毒 (rCV) 和重组濑户病毒 (rSEV)）的抗体，使我们能够确定这三种病毒之间的遗传和抗原关系。
The second open reading frame (ORF2) gene of the Chitta virus (CHV) was cloned to construct a recombinant baculovirus. The CHV ORF2 is predicted to encode a capsid protein of 535 amino acids (aa). CHV showed a high aa identity in the capsid region with genogroup II Norwalk virus (NV) (65–85%), but a low aa identity with genogroup I NV (44–46%). Phylogenetic analysis of the ORF2 gene demonstrated that CHV is genetically closely related to the Hawaii virus included in genogroup II NV. The recombinant capsid protein of CHV (rCHV) self‐assembled to form empty virus‐like particles (VLPs) when expressed in insect cells with the recombinant baculovirus. An enzyme‐linked immunosorbent assay (ELISA) based on antisera to rCHV was developed to detect CHV antigen in stools. The antigen ELISA appeared to be highly specific to both rCHV and CHV‐like strains. In addition, combined use of antigen ELISAs using antibodies against two antigenically distinct recombinant VLPs, the recombinant Chiba virus (rCV) and recombinant Seto virus (rSEV), enabled us to determine the genetic as well as antigenic relationship among these three viruses.
影响因子：
3.7
作者：
JIANG, X;WANG, M;ESTES, MK
通讯作者：
ESTES, MK