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重组杆状病毒表达的汉坦病毒和汉城病毒核衣壳蛋白的抗原特性:应用一种缺失两种病毒共有的抗原区域的截短蛋白作为血清分型抗原。

Antigenic characterization of Hantaan and Seoul virus nucleocapsid proteins expressed by recombinant baculovirus: application of a truncated protein, lacking an antigenic region common to the two viruses, as a serotyping antigen.

作者信息

Morii M, Yoshimatsu K, Arikawa J, Zhou G, Kariwa H, Takashima I

机构信息

Hokkaido University School of Medicine, Hokkaido University, Sapporo 060, Japan.

出版信息

J Clin Microbiol. 1998 Sep;36(9):2514-21. doi: 10.1128/JCM.36.9.2514-2521.1998.

DOI:10.1128/JCM.36.9.2514-2521.1998
PMID:9705385
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC105155/
Abstract

Hantaan virus (HTN) and Seoul virus (SEO) are members of the genus Hantavirus in the family Bunyaviridae and are causative agents of hemorrhagic fever with renal syndrome. The complete and truncated nucleocapsid proteins (NP) of HTN and SEO were expressed by a recombinant baculovirus system. Antigenic characterization of the NP using monoclonal antibodies (MAbs) indicated that the binding sites for the serotype-specific MAbs were located between amino acids (aa) 155 and 429. A Western blot assay indicated that the serotype-specific epitopes were conformation dependent. An indirect immunofluorescence antibody (IFA) assay with the truncated NP (aa 155 to 429) was able to distinguish convalescent-phase sera from HTN and SEO patients. However, the antibody titers with the truncated NP were lower than those with the whole NP. The truncated NP of SEO (aa 155 to 429) could be used as an enzyme-linked immunosorbent assay (ELISA) antigen, but the truncated NP from HTN lost its reactivity when used for ELISA. The IFA assay using baculovirus-expressed truncated NP as an antigen is a rapid, simple, and safe test for distinguishing between HTN and SEO infections by serotype.

摘要

汉坦病毒(HTN)和汉城病毒(SEO)是布尼亚病毒科汉坦病毒属的成员,是肾综合征出血热的病原体。通过重组杆状病毒系统表达了HTN和SEO的完整和截短核衣壳蛋白(NP)。使用单克隆抗体(MAb)对NP进行抗原特性分析表明,血清型特异性MAb的结合位点位于氨基酸(aa)155至429之间。蛋白质印迹分析表明,血清型特异性表位依赖于构象。用截短的NP(aa 155至429)进行间接免疫荧光抗体(IFA)检测能够区分HTN和SEO患者的恢复期血清。然而,截短NP的抗体滴度低于完整NP的抗体滴度。SEO的截短NP(aa 155至429)可用作酶联免疫吸附测定(ELISA)抗原,但HTN的截短NP用于ELISA时失去了反应性。使用杆状病毒表达的截短NP作为抗原的IFA检测是一种快速、简单且安全的通过血清型区分HTN和SEO感染的检测方法。

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