采用基于S蛋白的酶联免疫吸附测定（ELISA）和针对病毒刺突假型病毒的中和试验检测冠状病毒HKU1感染的血清流行率。

Examination of seroprevalence of coronavirus HKU1 infection with S protein-based ELISA and neutralization assay against viral spike pseudotyped virus.

作者信息

Chan C M, Tse Herman, Wong S S Y, Woo P C Y, Lau S K P, Chen L, Zheng B J, Huang J D, Yuen K Y

机构信息

State Key Laboratory of Emerging Infectious Diseases, Department of Microbiology, The University of Hong Kong, Hong Kong, PR China.

出版信息

J Clin Virol. 2009 May;45(1):54-60. doi: 10.1016/j.jcv.2009.02.011. Epub 2009 Apr 1.

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, from 0% in the age group of <10 years old to a plateau of 21.6% in the age group of 31-40 years old.

CONCLUSIONS

Our study demonstrated the usefulness of the S-based ELISA which could be applied to future epidemiological studies of HCoV-HKU1 in other localities.

背景

人冠状病毒HKU1（HCoV-HKU1）是一种最近发现的冠状病毒，在全球范围内均有分布，主要引起呼吸道感染。

目的

调查本地人群中HKU1感染的血清流行病学情况。

研究设计

开发了一种基于酶联免疫吸附测定（ELISA）的IgG抗体检测方法，使用重组HCoV-HKU1核衣壳蛋白和刺突（S）蛋白（A基因型）来诊断CoV-HKU1感染。此外，还开发了一种使用HCoV-HKU1假型病毒的中和抗体检测方法，以检测基于S蛋白的ELISA中抗体呈阳性的血清中中和抗体的存在。

结果

基于重组S蛋白的ELISA结果通过蛋白质印迹法、免疫荧光分析和流式细胞术进行了验证。联合结果显示与斯皮尔曼系数为0.94具有良好的相关性。在一家本地医院环境中使用这种新开发的ELISA进行的血清流行病学研究表明，HCoV-HKU1血清阳性率在儿童期和成年早期稳步上升，从<10岁年龄组的0%上升至31-40岁年龄组的21.6%的平稳水平。