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截短型汉坦病毒核衣壳蛋白的特性及其血清分型应用

Characterization of truncated hantavirus nucleocapsid proteins and their application for serotyping.

作者信息

Li Guangyu, Pan Lei, Mou Danlei, Chen Yanping, Zhang Yan, Li Xinhong, Ren Junping, Wang Pingzhong, Zhang Ying, Jia Zhansheng, Huang Changxing, Sun Yongtao, Yang Weisong, Xiao Shu-Yuan, Bai Xuefan

机构信息

Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.

出版信息

J Med Virol. 2006 Jul;78(7):926-32. doi: 10.1002/jmv.20643.

Abstract

Hemorrhagic fever with renal syndrome (HFRS) is a fulminant infectious disease characterized by fever, hemorrhage, renal impairment, and thrombocytopenia. Hantaviruses associated with this belong to different serotypes: Hantaan (HTN), Seoul (SEO), Dobrava/Belgrade (DOB), and Puumala (PUU). The first two, HTN and SEO, are endemic in China. To investigate the epidemiology of HFRS and virus transmission in China, we constructed prokaryotic plasmids encoding truncated recombinant HTN and SEO nucleocapsid proteins (NPs), which lacked 154 amino acid (aa), 99 aa, or 49 aa in the N-terminal region, respectively. After expression, the truncated rNPs were tested as serotyping antigens, particularly for use in the enzyme-linked immunosorbent assay (ELISA). In addition, 68 acute and 52 convalescent sera were collected from HFRS patients from Harbin, Lantian, and Kaifeng regions in China in 2004, which had hantavirus specific antibodies by IFA. A neutralization test was used to differentiate these, which showed that 73 were due to HTN infection, 33 to SEO infection, and 14 undetermined. By ELISA, the truncated rNPs, that lacked 99 (rNP100) or 49 (rNP50) N-terminal amino acids of the NPs of HTN and SEO, were able to differentiate HTNV and SEOV-specific immune sera, but the rNP155 could not. Particularly, the ELISAs based on the rNP50s had a result comparable to PRNT. Thus, the rNP50 is recommended as efficient serotyping antigen for hantavirus infection diagnosis by ELISA.

摘要

肾综合征出血热(HFRS)是一种以发热、出血、肾功能损害和血小板减少为特征的暴发性传染病。与之相关的汉坦病毒属于不同血清型:汉滩病毒(HTN)、汉城病毒(SEO)、多布拉伐/贝尔格莱德病毒(DOB)和普马拉病毒(PUU)。前两种,即HTN和SEO,在中国呈地方性流行。为了调查中国HFRS的流行病学及病毒传播情况,我们构建了编码截短的重组HTN和SEO核衣壳蛋白(NP)的原核质粒,这些截短的NP在N端区域分别缺失154个氨基酸(aa)、99个aa或49个aa。表达后,对截短的rNP进行了血清分型抗原测试,特别是用于酶联免疫吸附测定(ELISA)。此外,2004年从中国哈尔滨、蓝田和开封地区的HFRS患者中收集了68份急性期血清和52份恢复期血清,这些血清通过免疫荧光抗体法(IFA)检测具有汉坦病毒特异性抗体。采用中和试验对这些血清进行鉴别,结果显示73份由HTN感染引起,33份由SEO感染引起,14份未确定。通过ELISA检测,截短的rNP,即HTN和SEO的NP分别缺失99个(rNP100)或49个(rNP50)N端氨基酸,能够区分HTNV和SEOV特异性免疫血清,但rNP155不能。特别是,基于rNP50的ELISA结果与空斑减少中和试验(PRNT)相当。因此,推荐rNP50作为通过ELISA诊断汉坦病毒感染的有效血清分型抗原。

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