构建伪型人类冠状病毒并检测人群中已有的抗体

IF 3.5 Q1 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

Biosafety and Health Pub Date : 2024-10-01 DOI:10.1016/j.bsheal.2024.09.002

Qi Jiang , Xi Wu , Fangyu Dong , Shan Qiao , Qiaoyun Shi , Changyong Jian , Chen Chen , Jiuyue Zhou , Youchun Wang , Weijin Huang

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引用次数: 0

摘要

在 2019 年冠状病毒疾病（COVID-19）爆发前后，我们利用基于伪型病毒的冠状病毒抗体（Pseudotypes virus-based Coronavirus-Based抗体）对 HCoV、严重急性呼吸系统综合征（SARS）相关冠状病毒（SARS-CoV-1）、严重急性呼吸系统综合征冠状病毒 2（SARS-CoV-2）、中东呼吸系统综合征冠状病毒（MERS-CoV）、HCoV-229E、HCoV-NL63、HCoV-HKU1 和 HCoV-OC43 进行了检测。我们利用基于伪型病毒的中和试验（PBNA）或酶联免疫吸附试验（ELISA）检测了 2009-2010 年和 2023 年采集的血清样本中的 HCoV 抗体水平。PBNA 结果显示，SARS-CoV-1 和 MERS-CoV 的中和抗体呈阴性。在 2009 年至 2010 年的血清样本中，SARS-CoV-2（D614G）的中和抗体呈阴性，而在 2023 年的血清样本中，73 个样本（73%）与 SARS-CoV-2 D614G 株出现中和反应，96 个样本（96%）与 BA.5 株出现中和反应，91 个样本（91%）与 BF.7 株出现中和反应。在 COVID-19 前的样本中，33%（33/100）与 HCoV-229E 发生中和反应，63%（63/100）与 HCoV-NL63 发生中和反应。在 COVID-19 后的样本中，50%（50/100）的样本与 HCoV-229E 发生中和反应，49%（49/100）的样本与 HCoV-NL63 发生中和反应。由于α冠状病毒属的受体与其他β冠状病毒属不同，因此无法通过构建相应的伪型病毒来检测针对HCoV-OC43和HCoV-HKU1病毒的中和抗体。使用 ELISA 方法检测了针对 HCoV-OC43 和 HCoV-HKU1 病毒的结合抗体。结果显示，在 COVID-19 前的样本中，83%（83/100）和 45%（45/100）的样本与 HCoV-OC43 和 HCoV-HKU1 病毒具有结合活性。在 COVID-19 后的样本中，与 HCoV-OC43 和 HCoV-HKU1 的结合活性分别为 100%（100/100）和 81%（81/100）。

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Construction of pseudotyped human coronaviruses and detection of pre-existing antibodies in the human population

In order to clarify the pre-exist immunity background of different human coronaviruses (HCoV), this study investigated the positive rate of spike (S) protein antibodies of HCoV, including HCoV- severe acute respiratory syndrome (SARS) −associated coronavirus (SARS-CoV-1), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Middle East respiratory syndrome coronavirus (MERS-CoV), HCoV-229E, HCoV-NL63, HCoV-HKU1 and HCoV-OC43, before and after the Coronavirus Disease 2019 (COVID-19) outbreak. We utilized pseudotyped virus-based neutralization assays (PBNA) or enzyme-linked immunosorbent assays (ELISA) to detect antibody levels against HCoV in serum samples collected in 2009–2010 and 2023. The PBNA results showed that neutralizing antibodies against SARS-CoV-1 and the MERS-CoV were negative. In the serum samples from 2009 to 2010, neutralizing antibodies against SARS-CoV-2 (D614G) were negative, whereas in the serum samples from 2023, 73 samples (73 %) showed neutralizing reactions with the SARS-CoV-2 D614G strain, 96 samples (96 %) with the BA.5 strain, and 91 samples (91 %) with the BF.7 strain. Among pre-COVID-19 samples, 33 % (33/100) showed neutralizing reactions with HCoV-229E and 63 % (63/100) with HCoV-NL63. Among post-COVID-19 samples, 50 % (50/100) showed neutralizing reactions with HCoV-229E and 49 % (49/100) with HCoV-NL63. Due to the different receptors of alpha coronavirus genus compared to other beta coronavirus genus, neutralizing antibodies against HCoV-OC43 and HCoV-HKU1 virus cannot be detected by constructing corresponding pseudotyped virus. Binding antibodies against HCoV-OC43 and HCoV-HKU1 virus were detected using ELISA. The results revealed that among pre-COVID-19 samples, 83 % (83/100) and 45 % (45/100) had binding activity with HCoV-OC43 and HCoV-HKU1, respectively. Among post-COVID-19 samples, 100 % (100/100) and 81 % (81/100) had binding activity with HCoV-OC43 and HCoV-HKU1, respectively.

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