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新型桥梁多重 ELISA 法检测 SARS-CoV-2 抗体。

Novel bridge multi-species ELISA for detection of SARS-CoV-2 antibodies.

机构信息

Departamento de Microbiología, Inmunología, Biotecnología y Genética, Facultad de Farmacia y Bioquímica, Cátedra de Inmunología, Universidad de Buenos Aires, Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) Universidad de Buenos Aires, Instituto de Estudios de la Inmunidad Humoral "Prof. Ricardo A. Margni" (IDEHU), Buenos Aires, Argentina.

Departamento de Microbiología, Inmunología, Biotecnología y Genética, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Universidad de Buenos Aires, Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Buenos Aires, Argentina.

出版信息

J Immunol Methods. 2022 Dec;511:113365. doi: 10.1016/j.jim.2022.113365. Epub 2022 Oct 4.

Abstract

UNLABELLED

Considering the course of the current SARS-CoV-2 pandemic, it is important to have serological tests for monitoring humoral immune response against SARS-CoV-2 infection and vaccination. Herein we describe a novel bridge enzyme-linked immunosorbent assay (b-ELISA) for SARS-CoV-2 antibodies detection in human and other species, employing recombinant Spike protein as a unique antigen, which is produced at high scale in insect larvae.

METHODS

Eighty two human control sera/plasmas and 169 COVID-19 patients' sera/plasmas, confirmed by rRT-PCR, were analyzed by the b-ELISA assay. In addition, a total of 27 animal sera (5 horses, 13 rats, 2 cats and 7 dogs) were employed in order to evaluate the b-ELISA in other animal species.

RESULTS

Out of the 169 patient samples, 129 were positive for IgG anti-SARS-CoV-2 and 40 were negative when they were tested by ELISA COVIDAR® IgG. When a cut-off value of 5.0 SDs was established, 124 out of the 129 COVID-19 positive samples were also positive by our developed b-ELISA (sensitivity: 96.12%). Moreover, the test was able to evaluate the humoral immune response in animal models and also detected as positive a naturally infected cat and two dogs with symptoms, whose owners had suffered the COVID-19 disease.

CONCLUSION

The obtained results demonstrate that the method developed herein is versatile, as it is able to detect antibodies against SARS-CoV-2 in different animal species without the need to perform and optimize a new assay for each species.

摘要

目的

考虑到当前 SARS-CoV-2 大流行的进程,有必要进行血清学检测以监测针对 SARS-CoV-2 感染和疫苗接种的体液免疫反应。本研究旨在描述一种用于检测人类和其他物种 SARS-CoV-2 抗体的新型桥接酶联免疫吸附试验(b-ELISA),该方法使用重组 Spike 蛋白作为唯一抗原,该抗原可在昆虫幼虫中大规模生产。

方法

采用 b-ELISA 检测 82 份人类对照血清/血浆和 169 份经 rRT-PCR 确诊的 COVID-19 患者血清/血浆。此外,还使用了总共 27 份动物血清(5 匹马、13 只大鼠、2 只猫和 7 只狗)来评估该方法在其他动物物种中的应用。

结果

在 169 份患者样本中,有 129 份 IgG 抗 SARS-CoV-2 呈阳性,而 ELISA COVIDAR® IgG 检测有 40 份为阴性。当建立 5.0SD 作为截断值时,我们开发的 b-ELISA 检测出 124 份 COVID-19 阳性样本也呈阳性(敏感性:96.12%)。此外,该试验能够评估动物模型中的体液免疫反应,还能检测到患有 COVID-19 疾病的具有症状的自然感染猫和两只狗的抗体呈阳性。

结论

研究结果表明,所开发的方法具有通用性,因为它能够检测不同动物物种中针对 SARS-CoV-2 的抗体,而无需针对每种物种进行和优化新的检测方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab66/9529351/1fdf3f3ba1a8/gr1_lrg.jpg

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