Programa de Pós-Graduação Em Ciências da Saúde: Infectologia E Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, MG, 30.130-100, Brazil.
Centro de Tecnologia de Vacinas (CT Vacinas) / BH-Tec, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Sci Rep. 2023 Mar 16;13(1):4345. doi: 10.1038/s41598-023-31382-5.
Serological assays have been widely used to detect anti-SARS-CoV-2 antibodies, which are generated from previous exposure to the virus or after vaccination. The presence of anti-SARS-CoV-2 Nucleocapsid antibodies was recently reported in patients´ urine using an in-house urine-based ELISA-platform, allowing a non-invasive way to collect clinical samples and assess immune conversion. In the current study, we evaluated and validated another in-house urine-based ELISA for the detection of anti-SARS-CoV-2 Spike antibodies. Three partial recombinant SARS-CoV-2 Spike proteins comprising the Receptor Binding Domain, expressed in eukaryotic or prokaryotic systems, were tested in an ELISA platform against a panel of over 140 urine and paired serum samples collected from 106 patients confirmed positive for SARS-CoV-2 by qRT-PCR. The key findings from our study were that anti-SARS-CoV-2 Spike antibodies could be detected in urine samples and that the prokaryotic expression of the rSARS-CoV-2 Spike protein was not a barrier to obtain relatively high serology efficiency for the urine-based assay. Thus, use of a urine-based ELISA assay with partial rSARS-CoV-2 Spike proteins, expressed in a prokaryotic system, could be considered as a convenient tool for screening for the presence of anti-SARS-CoV-2 Spike antibodies, and overcome the difficulties arising from sample collection and the need for recombinant proteins produced with eukaryotic expression systems.
血清学检测已被广泛用于检测抗 SARS-CoV-2 抗体，这些抗体可由先前接触病毒或接种疫苗后产生。最近有研究报告称，使用基于尿液的内部 ELISA 平台在患者尿液中检测到抗 SARS-CoV-2 核衣壳抗体，这为非侵入性采集临床样本和评估免疫转化提供了可能。在本研究中，我们评估和验证了另一种基于尿液的内部 ELISA 平台，用于检测抗 SARS-CoV-2 刺突抗体。三个包含受体结合域的部分 SARS-CoV-2 刺突蛋白，分别在真核或原核系统中表达，在 ELISA 平台上针对 140 多个尿液样本和配对血清样本进行了测试，这些样本来自 106 名经 qRT-PCR 证实为 SARS-CoV-2 阳性的患者。我们研究的主要发现是，在尿液样本中可以检测到抗 SARS-CoV-2 刺突抗体，并且 rSARS-CoV-2 刺突蛋白的原核表达并不是获得基于尿液的检测相对较高血清学效率的障碍。因此，使用基于尿液的 ELISA 检测与部分 rSARS-CoV-2 刺突蛋白结合，该蛋白在原核系统中表达，可以被认为是一种方便的工具，用于筛选抗 SARS-CoV-2 刺突抗体的存在，并克服样本采集和需要使用真核表达系统产生重组蛋白带来的困难。
