从重组关键抗原到准确、可负担的血清学检测:从新冠疫情中汲取的经验教训,以应对未来大流行。

From a recombinant key antigen to an accurate, affordable serological test: Lessons learnt from COVID-19 for future pandemics.

作者信息

Alvim Renata G F, Lima Tulio M, Rodrigues Danielle A S, Marsili Federico F, Bozza Vicente B T, Higa Luiza M, Monteiro Fabio L, Abreu Daniel P B, Leitão Isabela C, Carvalho Renato S, Galliez Rafael M, Castineiras Terezinha M P P, Travassos Leonardo H, Nobrega Alberto, Tanuri Amilcar, Ferreira Orlando C, Vale André M, Castilho Leda R

机构信息

COPPE, Chemical Engineering Program, Cell Culture Engineering Lab, Universidade Federal do Rio de Janeiro - UFRJ, Brazil.

School of Chemistry, EPQB Program, Universidade Federal do Rio de Janeiro - UFRJ, Brazil.

出版信息

Biochem Eng J. 2022 Aug;186:108537. doi: 10.1016/j.bej.2022.108537. Epub 2022 Jul 16.

Abstract

Serological tests detect antibodies generated by infection or vaccination, and are indispensable tools along different phases of a pandemic, from early monitoring of pathogen spread up to seroepidemiological studies supporting immunization policies. This work discusses the development of an accurate and affordable COVID-19 antibody test, from production of a recombinant protein antigen up to test validation and economic analysis. We first developed a cost-effective, scalable technology to produce SARS-COV-2 spike protein and then used this antigen to develop an enzyme-linked immunosorbent assay (ELISA). A receiver operator characteristic (ROC) analysis allowed optimizing the cut-off and confirmed the high accuracy of the test: 98.6% specificity and 95% sensitivity for 11+ days after symptoms onset. We further showed that dried blood spots collected by finger pricking on simple test strips could replace conventional plasma/serum samples. A cost estimate was performed and revealed a final retail price in the range of one US dollar, reflecting the low cost of the ELISA test platform and the elimination of the need for venous blood sampling and refrigerated sample handling in clinical laboratories. The presented workflow can be completed in 4 months from first antigen expression to final test validation. It can be applied to other pathogens and in future pandemics, facilitating reliable and affordable seroepidemiological surveillance also in remote areas and in low-income countries.

摘要

血清学检测可检测由感染或疫苗接种产生的抗体,是大流行不同阶段不可或缺的工具,从病原体传播的早期监测到支持免疫政策的血清流行病学研究。这项工作讨论了一种准确且经济实惠的新冠病毒抗体检测方法的开发,从重组蛋白抗原的生产到检测验证和经济分析。我们首先开发了一种具有成本效益、可扩展的技术来生产新冠病毒刺突蛋白,然后使用该抗原来开发酶联免疫吸附测定(ELISA)。受试者工作特征(ROC)分析有助于优化临界值,并证实了该检测的高准确性:症状出现11天及以上后,特异性为98.6%,敏感性为95%。我们进一步表明,通过在简单试纸条上手指采血收集的干血斑可以替代传统的血浆/血清样本。进行了成本估算,结果显示最终零售价在1美元左右,这反映了ELISA检测平台的低成本,以及无需在临床实验室进行静脉采血和冷藏样本处理。从首次抗原表达至最终检测验证,所展示的工作流程可在4个月内完成。它可应用于其他病原体以及未来的大流行,也有助于在偏远地区和低收入国家进行可靠且经济实惠的血清流行病学监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b73e/9287463/10c95299bdc6/ga1_lrg.jpg

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