An indirect ELISA for detecting anti-SARS-CoV-2 antibodies in human sera using a baculovirus-expressed recombinant nucleocapsid antigen

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-05-01 DOI:10.1016/j.biologicals.2024.101769

Matheus Bernardes Torres Fogaça , Gildemar José Bezerra Crispim , Djairo Pastor Saavedra , Leonardo Lopes-Luz , Leonardo Assis da Silva , Brenda Rabello de Camargo , Rafael Alves Guimarães , Tatsuya Nagata , Bergmann Morais Ribeiro , Samira Bührer-Sékula

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引用次数: 0

Abstract

This study focuses on the development and initial assessment of an indirect IgG enzyme-linked immunosorbent assay (ELISA) specifically designed to detect of anti-SARS-CoV-2 antibodies. The unique aspect of this ELISA method lies in its utilization of a recombinant nucleocapsid (N) antigen, produced through baculovirus expression in insect cells. Our analysis involved 292 RT-qPCR confirmed positive serum samples and 54 pre-pandemic healthy controls. The process encompassed cloning, expression, and purification of the SARS-CoV-2 N gene in insect cells, with the resulted purified protein employed in our ELISA tests. Statistical analysis yielded an Area Under the Curve of 0.979, and the optimized cut-off exhibited 92 % sensitivity and 94 % specificity. These results highlight the ELISA's potential for robust and reliable serological detection of SARS-CoV-2 antibodies. Further assessments, including a larger panel size, reproducibility tests, and application in diverse populations, could enhance its utility as a valuable biotechnological solution for diseases surveillance.

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一种利用杆状病毒表达的重组核壳抗原检测人血清中抗 SARS-CoV-2 抗体的间接 ELISA 方法

本研究的重点是开发和初步评估一种间接 IgG 酶联免疫吸附试验（ELISA），专门用于检测抗 SARS-CoV-2 抗体。这种酶联免疫吸附试验方法的独特之处在于它使用了通过昆虫细胞中的杆状病毒表达产生的重组核壳（N）抗原。我们的分析涉及 292 份经 RT-qPCR 确认的阳性血清样本和 54 份大流行前健康对照样本。分析过程包括在昆虫细胞中克隆、表达和纯化 SARS-CoV-2 N 基因，并将纯化后的蛋白用于 ELISA 检测。统计分析得出的曲线下面积为 0.979，优化的临界值显示出 92% 的灵敏度和 94% 的特异性。这些结果凸显了该酶联免疫吸附试验在对 SARS-CoV-2 抗体进行稳健可靠的血清学检测方面的潜力。进一步的评估，包括更大的面板尺寸、重现性测试和在不同人群中的应用，可以提高其作为疾病监测的一种有价值的生物技术解决方案的实用性。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.