Examination of seroprevalence of coronavirus HKU1 infection with S protein-based ELISA and neutralization assay against viral spike pseudotyped virus
Examination of seroprevalence of coronavirus HKU1 infection with S protein-based ELISA and neutralization assay against viral spike pseudotyped virus
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DOI：
10.1016/j.jcv.2009.02.011
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发表时间：
2009-05-01
影响因子：
8.8
通讯作者：
Yuen, K. Y.
中科院分区：
文献类型：
--
作者：
Chan, C. M.;Tse, Herman;Yuen, K. Y.
背景：人类冠状病毒 HKU1 (HCoV-HKU1) 是最近发现的一种全球分布的冠状病毒，已知主要引起呼吸道感染。目的：调查当地人群中 HKU1 感染的血清流行病学。研究设计：开发了一种使用重组 HCoV-HKU1 核衣壳和刺突 (S) 蛋白（A 基因型）的基于 ELISA 的 IgG 抗体检测方法，用于诊断 CoV-HKU1 感染。开发了 HCoV-HKU1 假型病毒，用于在基于 S 蛋白的 ELISA 中检测血清中中和抗体的存在，并且抗体呈阳性。 结果：基于重组 S 蛋白的 ELISA 结果通过蛋白质印迹、免疫荧光分析和流式细胞术进行了验证。耦合结果表明与 Spearmen 系数 0.94 具有良好的相关性。使用这种新开发的 ELISA 在当地医院进行的血清流行病学研究显示，儿童和成年早期的 HCoV-HKU1 血清流行率稳步上升，在 20 岁以下年龄组中前 0%
Background: Human coronavirus HKU1 (HCoV-HKU1) is a recently identified coronavirus with a global distribution and known to cause mainly respiratory infections.Objectives: To investigate the seroepidemiology of HKU1 infections in our local population.Study design: An ELISA-based IgG antibody detection assay using recombinant HCoV-HKU1 nucleocapsid and spike (S) proteins (genotype A) were developed for the diagnosis of CoV-HKU1 infections, Additionally, a neutralization antibody assay using the HCoV-HKU1 pseudotyped virus was developed to detect the presence of neutralizing antibodies in serum with antibody positivity in an S protein-based ELISA.Results: Results of the recombinant S protein-based ELISA were validated with Western blot, immunofluorescence analysis and flow cytometry. The coupled results demonstrated good correlation with Spearmen's coefficient of 0.94. Seroepidemiological study in a local hospital-based setting using this newly developed ELISA showed steadily increasing HCoV-HKU1 seroprevalence in childhood and early adulthood, front 0% in the age group of