期刊
VACCINES
卷 9, 期 1, 页码 -出版社
MDPI
DOI: 10.3390/vaccines9010035
关键词
SARS-CoV-2; Covid-19; serological test; epitope mapping; peptide microarrays; click chemistry
资金
- Regione Lombardia, project READY (Regional Network for developing diagnostic methods in rapid response to emerging epidemics and bio-emergencies) [229472]
- European Union [874735]
- Italian Ministry of Health Fondi Ricerca corrente-L1P5
A workflow for rapid epitope discovery of SARS-CoV-2 on peptide microarrays was reported, leading to the identification of immunodominant regions on the spike, nucleocapsid, and Orf1ab proteins. The epitope of N protein's region 155-171 showed good diagnostic performance in discriminating Covid-19 positive individuals, with 92% sensitivity and 100% specificity for IgG detection. This epitope represents a promising candidate for further development and rapid implementation in serological tests.
A workflow for rapid SARS-CoV-2 epitope discovery on peptide microarrays is herein reported. The process started with a proteome-wide screening of immunoreactivity based on the use of a high-density microarray followed by a refinement and validation phase on a restricted panel of probes using microarrays with tailored peptide immobilization through a click-based strategy. Progressively larger, independent cohorts of Covid-19 positive sera were tested in the refinement processes, leading to the identification of immunodominant regions on SARS-CoV-2 spike (S), nucleocapsid (N) protein and Orf1ab polyprotein. A summary study testing 50 serum samples highlighted an epitope of the N protein (region 155-71) providing good diagnostic performance in discriminating Covid-19 positive vs. healthy individuals. Using this epitope, 92% sensitivity and 100% specificity were reached for IgG detection in Covid-19 samples, and no cross-reactivity with common cold coronaviruses was detected. Likewise, IgM immunoreactivity in samples collected within the first month after symptoms onset showed discrimination ability. Overall, epitope 155-171 from N protein represents a promising candidate for further development and rapid implementation in serological tests.
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