tetano
Editor, Senior Moderator
Eur J Clin Microbiol Infect Dis
. 2020 Nov 12.
doi: 10.1007/s10096-020-04102-4. Online ahead of print.
Development and evaluation of a serological test for diagnosis of COVID-19 with selected recombinant spike proteins
Pingping Zhang[SUP] 1 2 [/SUP], Qi Gao[SUP] 2 3 [/SUP], Tang Wang[SUP] 4 [/SUP], Yuehua Ke[SUP] 5 [/SUP], Fei Mo[SUP] 6 7 [/SUP], Ruizhong Jia[SUP] 5 [/SUP], Wanbing Liu[SUP] 8 [/SUP], Lei Liu[SUP] 8 [/SUP], Shangen Zheng[SUP] 8 [/SUP], Yuzhen Liu[SUP] 9 [/SUP], Luping Li[SUP] 10 [/SUP], Yao Wang[SUP] 11 [/SUP], Lei Xu[SUP] 2 3 [/SUP], Kun Hao[SUP] 2 3 [/SUP], Wei Min[SUP] 2 3 [/SUP], Xiaoli Liu[SUP] 2 3 [/SUP], Ruifu Yang[SUP] 1 2 [/SUP], Shiyue Li[SUP] 12 [/SUP], Changqing Lin[SUP] 13 14 [/SUP], Yong Zhao[SUP] 15 16 [/SUP]
Affiliations
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: Colloidal gold immunochromatography assay (GICA); Recombinant protein; SARS-CoV-2; Serological diagnosis.
. 2020 Nov 12.
doi: 10.1007/s10096-020-04102-4. Online ahead of print.
Development and evaluation of a serological test for diagnosis of COVID-19 with selected recombinant spike proteins
Pingping Zhang[SUP] 1 2 [/SUP], Qi Gao[SUP] 2 3 [/SUP], Tang Wang[SUP] 4 [/SUP], Yuehua Ke[SUP] 5 [/SUP], Fei Mo[SUP] 6 7 [/SUP], Ruizhong Jia[SUP] 5 [/SUP], Wanbing Liu[SUP] 8 [/SUP], Lei Liu[SUP] 8 [/SUP], Shangen Zheng[SUP] 8 [/SUP], Yuzhen Liu[SUP] 9 [/SUP], Luping Li[SUP] 10 [/SUP], Yao Wang[SUP] 11 [/SUP], Lei Xu[SUP] 2 3 [/SUP], Kun Hao[SUP] 2 3 [/SUP], Wei Min[SUP] 2 3 [/SUP], Xiaoli Liu[SUP] 2 3 [/SUP], Ruifu Yang[SUP] 1 2 [/SUP], Shiyue Li[SUP] 12 [/SUP], Changqing Lin[SUP] 13 14 [/SUP], Yong Zhao[SUP] 15 16 [/SUP]
Affiliations
- PMID: 33184753
- DOI: 10.1007/s10096-020-04102-4
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: Colloidal gold immunochromatography assay (GICA); Recombinant protein; SARS-CoV-2; Serological diagnosis.