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比较分析来自人类血清样本的 SARS-CoV-2 结合抗体(IgG、IgM、IgA)和中和抗体。

Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples.

机构信息

VisMederi S.r.l., Siena, Italy.

VisMederi Research S.r.l., Siena, Italy.

出版信息

J Immunol Methods. 2021 Feb;489:112937. doi: 10.1016/j.jim.2020.112937. Epub 2020 Nov 28.

Abstract

A newly identified coronavirus, named SARS-CoV-2, emerged in December 2019 in Hubei Province, China, and quickly spread throughout the world; so far, it has caused more than 49.7 million cases of disease and 1,2 million deaths. The diagnosis of SARS-CoV-2 infection is currently based on the detection of viral RNA in nasopharyngeal swabs by means of molecular-based assays, such as real-time RT-PCR. Furthermore, serological assays detecting different classes of antibodies constitute an excellent surveillance strategy for gathering information on the humoral immune response to infection and the spread of the virus through the population. In addition, it can contribute to evaluate the immunogenicity of novel future vaccines and medicines for the treatment and prevention of COVID-19 disease. The aim of this study was to determine SARS-CoV-2-specific antibodies in human serum samples by means of different commercial and in-house ELISA kits, in order to evaluate and compare their results first with one another and then with those yielded by functional assays using wild-type virus. It is important to identify the level of SARS-CoV-2-specific IgM, IgG and IgA antibodies in order to predict human population immunity, possible cross-reactivity with other coronaviruses and to identify potentially infectious subjects. In addition, in a small sub-group of samples, a subtyping IgG ELISA has been performed. Our findings showed a notable statistical correlation between the neutralization titers and the IgG, IgM and IgA ELISA responses against the receptor-binding domain of the spike protein. Thus confirming that antibodies against this portion of the virus spike protein are highly neutralizing and that the ELISA Receptor-Binding Domain-based assay can be used as a valid surrogate for the neutralization assay in laboratories that do not have biosecurity level-3 facilities.

摘要

一种新鉴定的冠状病毒,命名为 SARS-CoV-2,于 2019 年 12 月在中国湖北省出现,迅速在全球范围内传播;迄今为止,它已导致超过 4970 万例疾病和 120 万人死亡。SARS-CoV-2 感染的诊断目前基于通过分子检测法,如实时 RT-PCR,检测鼻咽拭子中的病毒 RNA。此外,检测不同类别抗体的血清学检测构成了一种极好的监测策略,可用于收集有关感染和病毒在人群中传播的体液免疫反应的信息。此外,它有助于评估新型未来疫苗和治疗 COVID-19 疾病的药物的免疫原性。本研究的目的是通过不同的商业和内部 ELISA 试剂盒来确定人类血清样本中的 SARS-CoV-2 特异性抗体,以便首先评估和比较它们彼此之间的结果,然后与使用野生型病毒的功能测定的结果进行比较。确定 SARS-CoV-2 特异性 IgM、IgG 和 IgA 抗体的水平对于预测人群免疫力、与其他冠状病毒的可能交叉反应以及识别潜在的传染性个体非常重要。此外,在一小部分样本中,进行了亚类 IgG ELISA。我们的研究结果表明,中和滴度与针对刺突蛋白受体结合域的 IgG、IgM 和 IgA ELISA 反应之间存在显著的统计学相关性。这证实了针对该病毒刺突蛋白这一部分的抗体具有高度中和作用,并且基于 ELISA 受体结合域的测定可以作为没有生物安全 3 级设施的实验室的中和测定的有效替代物。

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