Development of a Nucleocapsid-Based Human Coronavirus Immunoassay and Estimates of Individuals Exposed to Coronavirus in a US Metropolitan Population
Development of a Nucleocapsid-Based Human Coronavirus Immunoassay and Estimates of Individuals Exposed to Coronavirus in a US Metropolitan Population
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DOI：
10.1128/cvi.00124-08
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发表时间：
2008-12-01
影响因子：
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通讯作者：
Viscidi, Raphael P.
中科院分区：
文献类型：
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作者：
Severance, Emily G.;Bossis, Ioannis;Viscidi, Raphael P.
冠状病毒引起的呼吸道感染范围从普通感冒到严重急性呼吸道综合征（SARS）。非SARS冠状病毒暴露的估计值很高，特别是229 E和OC 43;然而，关于HKU 1和NL 63血清阳性率的信息较少。为了测量这四种冠状病毒株（229 E、HKU 1、NL 63和OC 43）的暴露率，我们设计了一种基于氨基和羧基末端标记的重组冠状病毒核衣壳抗原的免疫测定法。将四种人和一种猫冠状病毒抗原克隆到昆虫细胞中表达的杆状病毒中，并在基于酶联免疫吸附测定的系统的固相中回收结合的蛋白质。我们筛选了10名儿童和196名成人的血清，并根据主要血清阴性儿童的免疫球蛋白G（IgG）抗体水平建立了主要的临界点。每种冠状病毒的血清阳性成人比例如下：229 E，91.3%; HKU 1，59.2%; NL 63，91.8%;和OC 43，90.8%。未观察到对猫冠状病毒的显著血清学反应的证据。冠状病毒血清阳性和抗体水平与年龄，性别，种族，社会经济状况，吸烟状况和抽血季节的显着关联进行了卡方和回归分析。组II冠状病毒（OC 43和HKU 1）与种族显著相关（P
Coronaviruses cause respiratory infections ranging from common colds to severe acute respiratory syndrome (SARS) in humans. Estimates for exposure to non-SARS coronaviruses are high, particularly for 229E and OC43; however, less information regarding seroprevalence is available for HKU1 and NL63. To measure exposure rates to these four coronavirus strains (229E, HKU1, NL63, and OC43), we devised an immunoassay based on amino-and carboxy-terminally tagged recombinant coronavirus nucleocapsid antigens. Four human and one feline coronavirus antigen were cloned into baculoviruses expressed in insect cells and recovered proteins bound in the solid phase of an enzyme-linked immunosorbent assay-based system. We screened sera from 10 children and 196 adults and established primary cutoff points based on immunoglobulin G (IgG) antibody levels of the predominantly seronegative children. The proportion of seropositive adults for each coronavirus was as follows: 229E, 91.3%; HKU1, 59.2%; NL63, 91.8%; and OC43, 90.8%. No evidence of a significant serological response to the feline coronavirus was observed. Significant associations of coronavirus seropositivity and antibody levels with age, gender, race, socioeconomic status, smoking status, and season of the blood draw were tested with chi-square and regression analyses. The group II coronaviruses (OC43 and HKU1) were significantly associated with race (P