Development and evaluation of a serological test for diagnosis of COVID-19 with selected recombinant spike proteins
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ORIGINAL ARTICLE
Development and evaluation of a serological test for diagnosis of COVID-19 with selected recombinant spike proteins Pingping Zhang 1,2 & Qi Gao 2,3 & Tang Wang 4 & Yuehua Ke 5 & Fei Mo 6,7 & Ruizhong Jia 5 & Wanbing Liu 8 & Lei Liu 8 & Shangen Zheng 8 & Yuzhen Liu 9 & Luping Li 10 & Yao Wang 11 & Lei Xu 2,3 & Kun Hao 2,3 & Wei Min 2,3 & Xiaoli Liu 2,3 & Ruifu Yang 1,2 & Shiyue Li 4 & Changqing Lin 2,3 & Yong Zhao 1,2 Received: 13 July 2020 / Accepted: 5 November 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract 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. Keywords SARS-CoV-2 . Recombinant protein . Serological diagnosis . Colloidal gold immunochromatography assay (GICA)
Pingping Zhang, Qi Gao and Tang Wang contributed equally to this work. Ruifu Yang is a senior author * Shiyue Li [email protected] * Changqing Lin [email protected]
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Huoshenshan Hospital, Wuhan 430101, People’s Republic of China
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Center for Clinical Laboratories, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, People’s Republic of China
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Department of Basic Clinical Laboratory Medicine, School of Clinical Laboratory Science, Guizhou Medical University, Guiyang 550004, People’s Republic of China
* Yong Zhao [email protected] 8
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, People’s Republic of China
General Hospital of Central Theater Command of the PLA, Wuhan 430070, People’s Republic of China
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Beijing Key Laboratory of POCT for Bioemergency and Clinic, Beijing 100071, People’s Republic of China
Shijiazhuang Fifth Hospital, Shijiazhuang 050023, People’s Republic of China
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Beijing Hotgen Biotechnology Inc., Beijing 102600, People’s Republic
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