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通过定量抗原捕获酶联免疫吸附测定法确定感染细胞中博尔纳病病毒-p40和-p24蛋白之间的分子比率。

Molecular ratio between borna disease viral-p40 and -p24 proteins in infected cells determined by quantitative antigen capture ELISA.

作者信息

Watanabe M, Zhong Q, Kobayashi T, Kamitani W, Tomonaga K, Ikuta K

机构信息

Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.

出版信息

Microbiol Immunol. 2000;44(9):765-72. doi: 10.1111/j.1348-0421.2000.tb02561.x.

Abstract

We developed the antigen capture enzyme-linked immunosorbent assay (ELISA) systems for quantification of Borna disease virus (BDV) major antigens, p40 and p24. Using these ELISAs, we quantified the two proteins in various BDV-infected materials, including the cell lysates and culture supernatants as well as the homogenates of experimental animal brains. The ELISAs were also applied to measure the infectious titer of BDV in persistently infected cell lines. Quantitative analysis with these ELISAs allowed us to measure the molecular ratio between the p40 and p24 in infected samples. Interestingly, the ratio of p24 to p40 in persistently infected cells was much higher than that observed in acutely infected cells although the ratios in the supernatants from both cell lines were quite similar. BDV-inoculated gerbil brain cells showed a relatively high ratio of p24 to p40 as compared with acutely infected cells. These observations suggested that the molecular ratio between the proteins strongly depended on the infectious status of BDV in the host cells. The ELISA system developed here could be a convenient method for the quantification of BDV infection and may also be beneficial for understanding viral replication and infectious status in the host cells.

摘要

我们开发了用于定量博尔纳病病毒(BDV)主要抗原p40和p24的抗原捕获酶联免疫吸附测定(ELISA)系统。使用这些ELISA,我们对各种BDV感染材料中的这两种蛋白质进行了定量,包括细胞裂解物、培养上清液以及实验动物脑匀浆。这些ELISA还用于测量持续感染细胞系中BDV的感染滴度。通过这些ELISA进行的定量分析使我们能够测量感染样品中p40和p24之间的分子比例。有趣的是,尽管两种细胞系上清液中的比例相当相似,但持续感染细胞中p24与p40的比例远高于急性感染细胞中的比例。与急性感染细胞相比,接种BDV的沙鼠脑细胞显示出相对较高的p24与p40比例。这些观察结果表明,这些蛋白质之间的分子比例强烈依赖于BDV在宿主细胞中的感染状态。这里开发的ELISA系统可能是一种用于定量BDV感染的便捷方法,也可能有助于了解病毒在宿主细胞中的复制和感染状态。

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