Sánchez Belén G, Bort Alicia, Mora-Rodríguez José María, Díaz-Yuste Alba, Gasalla José Manuel, Sánchez-Chapado Manuel, Sebastián-Martín Alba, Díaz-Laviada Inés
Biochemistry and Molecular Biology Unit, Department of Systems Biology, School of Medicine and Health Sciences, University of Alcalá, 28871 Alcalá de Henares, Madrid, Spain.
Department of Comparative Medicine, School of Medicine, Yale University, New Haven, CT 06519, USA.
Biomedicines. 2022 Nov 11;10(11):2897. doi: 10.3390/biomedicines10112897.
Throughout the pandemic, serological assays have been revealed as crucial for detecting previous exposures to the virus and determining the timing of antibody maintenance after vaccination or natural infection. This study aimed to develop an optimized enzyme-linked immunosorbent assay (ELISA)-based serology, which could be used in case of reagent shortages, such as that occurred in the beginning of this health emergency. As a result, we present a high-sensitive immunoassay for the determination of IgG levels in venous serum samples, using 2 μg/mL antigen (receptor-binding domain of the spike protein S1) for coating the plate and utilizing human samples at a dilution 1:1000. This method showed non-inferiority features versus a commercial kit, is less expensive, and has a higher spectrophotometric range that allows for a better quantification of the antibody titers. The optical density values before and after heating venous serum samples at 56 °C during 30 min was quite similar, showing that heat inactivation can be used to reduce the biohazardous risks while handling samples. Furthermore, we show that finger-stick capillary blood samples can also serve as a suitable source for IgG detection, bypassing the need for serum isolation and being suitable for point-of-care application (Pearson's coefficient correlation with capillary serum was 0.95, being statistically significant).
在整个疫情期间,血清学检测已被证明对于检测既往病毒暴露情况以及确定疫苗接种或自然感染后抗体维持的时间至关重要。本研究旨在开发一种基于酶联免疫吸附测定(ELISA)的优化血清学检测方法,该方法可在试剂短缺的情况下使用,比如在此次卫生紧急事件初期出现的试剂短缺情况。结果,我们展示了一种用于测定静脉血清样本中IgG水平的高灵敏度免疫测定方法,使用2μg/mL抗原(刺突蛋白S1的受体结合域)包被酶标板,并将人类样本稀释1:1000使用。该方法与商业试剂盒相比具有非劣效性,成本更低,并且具有更高的分光光度范围,能够更好地定量抗体滴度。在56℃加热静脉血清样本30分钟前后的光密度值非常相似,表明热灭活可用于在处理样本时降低生物危害风险。此外,我们表明指尖毛细血管血样本也可作为IgG检测的合适来源,无需进行血清分离,适用于即时检测应用(与毛细血管血清的皮尔逊系数相关性为0.95,具有统计学意义)。