SARS-CoV-2 Antigens Expressed in Plants Detect Antibody Responses in COVID-19 Patients
Copyright © 2021 Makatsa, Tincho, Wendoh, Ismail, Nesamari, Pera, de Beer, David, Jugwanth, Gededzha, Mampeule, Sanne, Stevens, Scott, Blackburn, Mayne, Keeton and Burgers..
Background: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has swept the world and poses a significant global threat to lives and livelihoods, with 115 million confirmed cases and at least 2.5 million deaths from Coronavirus disease 2019 (COVID-19) in the first year of the pandemic. Developing tools to measure seroprevalence and understand protective immunity to SARS-CoV-2 is a priority. We aimed to develop a serological assay using plant-derived recombinant viral proteins, which represent important tools in less-resourced settings. Methods: We established an indirect ELISA using the S1 and receptor-binding domain (RBD) portions of the spike protein from SARS-CoV-2, expressed in Nicotiana benthamiana. We measured antibody responses in sera from South African patients (n = 77) who had tested positive by PCR for SARS-CoV-2. Samples were taken a median of 6 weeks after the diagnosis, and the majority of participants had mild and moderate COVID-19 disease. In addition, we tested the reactivity of pre-pandemic plasma (n = 58) and compared the performance of our in-house ELISA with a commercial assay. We also determined whether our assay could detect SARS-CoV-2-specific IgG and IgA in saliva. Results: We demonstrate that SARS-CoV-2-specific immunoglobulins are readily detectable using recombinant plant-derived viral proteins, in patients who tested positive for SARS-CoV-2 by PCR. Reactivity to S1 and RBD was detected in 51 (66%) and 48 (62%) of participants, respectively. Notably, we detected 100% of samples identified as having S1-specific antibodies by a validated, high sensitivity commercial ELISA, and optical density (OD) values were strongly and significantly correlated between the two assays. For the pre-pandemic plasma, 1/58 (1.7%) of samples were positive, indicating a high specificity for SARS-CoV-2 in our ELISA. SARS-CoV-2-specific IgG correlated significantly with IgA and IgM responses. Endpoint titers of S1- and RBD-specific immunoglobulins ranged from 1:50 to 1:3,200. S1-specific IgG and IgA were found in saliva samples from convalescent volunteers. Conclusion: We demonstrate that recombinant SARS-CoV-2 proteins produced in plants enable robust detection of SARS-CoV-2 humoral responses. This assay can be used for seroepidemiological studies and to measure the strength and durability of antibody responses to SARS-CoV-2 in infected patients in our setting.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
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| Enthalten in: |
Frontiers in plant science - 12(2021) vom: 16., Seite 589940 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Makatsa, Mohau S [VerfasserIn] |
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| Links: |
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| Themen: |
COVID-19 |
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| Anmerkungen: |
Date Revised 11.02.2024 published: Electronic-eCollection Citation Status PubMed-not-MEDLINE |
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| doi: |
10.3389/fpls.2021.589940 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a Background: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has swept the world and poses a significant global threat to lives and livelihoods, with 115 million confirmed cases and at least 2.5 million deaths from Coronavirus disease 2019 (COVID-19) in the first year of the pandemic. Developing tools to measure seroprevalence and understand protective immunity to SARS-CoV-2 is a priority. We aimed to develop a serological assay using plant-derived recombinant viral proteins, which represent important tools in less-resourced settings. Methods: We established an indirect ELISA using the S1 and receptor-binding domain (RBD) portions of the spike protein from SARS-CoV-2, expressed in Nicotiana benthamiana. We measured antibody responses in sera from South African patients (n = 77) who had tested positive by PCR for SARS-CoV-2. Samples were taken a median of 6 weeks after the diagnosis, and the majority of participants had mild and moderate COVID-19 disease. In addition, we tested the reactivity of pre-pandemic plasma (n = 58) and compared the performance of our in-house ELISA with a commercial assay. We also determined whether our assay could detect SARS-CoV-2-specific IgG and IgA in saliva. Results: We demonstrate that SARS-CoV-2-specific immunoglobulins are readily detectable using recombinant plant-derived viral proteins, in patients who tested positive for SARS-CoV-2 by PCR. Reactivity to S1 and RBD was detected in 51 (66%) and 48 (62%) of participants, respectively. Notably, we detected 100% of samples identified as having S1-specific antibodies by a validated, high sensitivity commercial ELISA, and optical density (OD) values were strongly and significantly correlated between the two assays. For the pre-pandemic plasma, 1/58 (1.7%) of samples were positive, indicating a high specificity for SARS-CoV-2 in our ELISA. SARS-CoV-2-specific IgG correlated significantly with IgA and IgM responses. Endpoint titers of S1- and RBD-specific immunoglobulins ranged from 1:50 to 1:3,200. S1-specific IgG and IgA were found in saliva samples from convalescent volunteers. Conclusion: We demonstrate that recombinant SARS-CoV-2 proteins produced in plants enable robust detection of SARS-CoV-2 humoral responses. This assay can be used for seroepidemiological studies and to measure the strength and durability of antibody responses to SARS-CoV-2 in infected patients in our setting | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a COVID-19 | |
| 650 | 4 | |a ELISA | |
| 650 | 4 | |a SARS-CoV-2 | |
| 650 | 4 | |a plant expression | |
| 650 | 4 | |a serology | |
| 700 | 1 | |a Tincho, Marius B |e verfasserin |4 aut | |
| 700 | 1 | |a Wendoh, Jerome M |e verfasserin |4 aut | |
| 700 | 1 | |a Ismail, Sherazaan D |e verfasserin |4 aut | |
| 700 | 1 | |a Nesamari, Rofhiwa |e verfasserin |4 aut | |
| 700 | 1 | |a Pera, Francisco |e verfasserin |4 aut | |
| 700 | 1 | |a de Beer, Scott |e verfasserin |4 aut | |
| 700 | 1 | |a David, Anura |e verfasserin |4 aut | |
| 700 | 1 | |a Jugwanth, Sarika |e verfasserin |4 aut | |
| 700 | 1 | |a Gededzha, Maemu P |e verfasserin |4 aut | |
| 700 | 1 | |a Mampeule, Nakampe |e verfasserin |4 aut | |
| 700 | 1 | |a Sanne, Ian |e verfasserin |4 aut | |
| 700 | 1 | |a Stevens, Wendy |e verfasserin |4 aut | |
| 700 | 1 | |a Scott, Lesley |e verfasserin |4 aut | |
| 700 | 1 | |a Blackburn, Jonathan |e verfasserin |4 aut | |
| 700 | 1 | |a Mayne, Elizabeth S |e verfasserin |4 aut | |
| 700 | 1 | |a Keeton, Roanne S |e verfasserin |4 aut | |
| 700 | 1 | |a Burgers, Wendy A |e verfasserin |4 aut | |
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