Chávez-Valdés Sheila, Marichal-Rodríguez Ana K, Chacón-Quintero Yahima, Martínez-Rosales Ricardo, Gómez-Hernández Nivaldo, Ávila-Díaz Lismary, Vázquez-Arteaga Amalia, González-Formental Hany, Freyre-Corrales Giselle, Coizeau-Rodríguez Edelgis, Guillen Gerardo, Lemos-Pérez Gilda
Center for Genetic Engineering and Biotechnology, CIGB, Ave. 31 E/ 158 y 190, P.O. Box. 6162, La Habana, 10600, Cuba.
Center for Genetic Engineering and Biotechnology, CIGB, Ave. 31 E/ 158 y 190, P.O. Box. 6162, La Habana, 10600, Cuba; Latin American School of Medicine (ELAM), Carretera Panamericana Km 3 1/2, Carr. Panamericana, La Habana, 19108, Cuba.
Anal Biochem. 2025 May;700:115787. doi: 10.1016/j.ab.2025.115787. Epub 2025 Jan 31.
COVID-19 is caused by SARS-CoV-2, first identified in 2019. The Cuban vaccines, Abdala and Mambisa, have demonstrated efficacy in preventing SARS-CoV-2 infection. Immunoglobulin A (IgA) are the main line of defense against pathogens invading the respiratory or digestive tract and its presence in serum can be measured to assess vaccine efficacy. ELISAs are a valuable tool for assessing vaccine immunogenicity. These tests should be validated to ensure their reliability and suitability. The objective of this study was to validate a non-commercial ELISA for the quantification of total anti-RBD IgA in serum samples to support clinical studies. This assay demonstrated high clinical specificity (97.3 %). The accuracy and precision of the assay showed an overall error of less than 20 % at all levels in QCs. Re-evaluation of samples showed a mean difference of less than 30 % in 90.2 % of cases. Anti-RBD IgA titers correlated with viral neutralization titers and percentage inhibition of RBD-ACE2 binding. This assay was found to be highly accurate and reproducible for the quantification of anti-RBD IgA, met the most stringent acceptance criteria and is fit for purpose. It is currently being used to evaluate the immunogenicity of the Abdala and Mambisa vaccines.
新冠病毒病(COVID-19)由严重急性呼吸综合征冠状病毒2(SARS-CoV-2)引起,该病毒于2019年首次被发现。古巴的阿卜杜拉(Abdala)和曼比萨(Mambisa)疫苗已证明在预防SARS-CoV-2感染方面具有疗效。免疫球蛋白A(IgA)是抵御侵入呼吸道或消化道病原体的主要防线,可以通过检测其在血清中的存在情况来评估疫苗疗效。酶联免疫吸附测定(ELISA)是评估疫苗免疫原性的重要工具。这些检测应经过验证以确保其可靠性和适用性。本研究的目的是验证一种用于定量血清样本中总抗受体结合域(RBD)IgA的非商业ELISA方法,以支持临床研究。该检测方法显示出较高的临床特异性(97.3%)。该检测方法的准确性和精密度表明,在所有质量控制(QC)水平下,总体误差均小于20%。对样本的重新评估显示,90.2%的病例平均差异小于30%。抗RBD IgA滴度与病毒中和滴度以及RBD-血管紧张素转换酶2(ACE2)结合抑制百分比相关。结果发现,该检测方法在定量抗RBD IgA方面具有高度准确性和可重复性,符合最严格的验收标准且适用。目前该方法正用于评估阿卜杜拉和曼比萨疫苗的免疫原性。