Author: Zhang, Pingping; Gao, Qi; Wang, Tang; Ke, Yuehua; Mo, Fei; Jia, Ruizhong; Liu, Wanbing; Liu, Lei; Zheng, Shangen; Liu, Yuzhen; Li, Luping; Wang, Yao; Xu, Lei; Hao, Kun; Min, Wei; Liu, Xiaoli; Yang, Ruifu; Li, Shiyue; Lin, Changqing; Zhao, Yong
Title: Development and evaluation of a serological test for diagnosis of COVID-19 with selected recombinant spike proteins Cord-id: d9qb9zaa Document date: 2020_11_12
ID: d9qb9zaa
Snippet: Serological test is a valuable diagnostic tool for coronavirus disease 2019 (COVID-19). However, considerable improvements to these tests are needed, especially in the detection sensitivity. In this study, six recombinant nucleocapsid and spike proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were prepared and evaluated, including three prokaryotic expression nucleocapsid proteins (rN, rN1, rN2) and three eukaryotic expression spike proteins (rS1, rS-RBD, rS-RBD-mFc). The
Document: Serological test is a valuable diagnostic tool for coronavirus disease 2019 (COVID-19). However, considerable improvements to these tests are needed, especially in the detection sensitivity. In this study, six recombinant nucleocapsid and spike proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were prepared and evaluated, including three prokaryotic expression nucleocapsid proteins (rN, rN1, rN2) and three eukaryotic expression spike proteins (rS1, rS-RBD, rS-RBD-mFc). The recombinant proteins with the highest ELISA titers (rS1 and rS-RBD-mFc) were selected to develop a double-antigen sandwich colloidal gold immunochromatography assay (GICA) to detect total antibodies against SARS-CoV-2. The clinical evaluation results showed that the sensitivity and specificity of GICA were 92.09% (419/455) and 99.44% (706/710), respectively. Moreover, a significant number (65.63%, 21/32) of COVID-19 patients with undetectable viral RNA were correctly diagnosed by the GICA method. In conclusion, the eukaryotic expression spike proteins (rS1 and rS-RBD-mFc) are more suitable than the prokaryotic expression nucleocapsid proteins for serological diagnosis of SARS-CoV-2. The proposed GICA for detection of total antibodies could be a powerful complement to the current RNA tests for COVID-19. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10096-020-04102-4.
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