A novel highly specific biotinylated MAC-ELISA for detection of anti-SARS-CoV-2 nucleocapsid antigen IgM antibodies during the acute phase of COVID-19
© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia..
The gold standard for diagnosing COVID-19 in the acute phase is RT-qPCR. However, this molecular technique can yield false-negative results when nasopharyngeal swab collection is not conducted during viremia. To mitigate this challenge, the enzyme-linked immunosorbent assay (ELISA) identifies anti-SARS-CoV-2 IgM antibodies in the initial weeks after symptom onset, facilitating early COVID-19 diagnosis. This study introduces a novel and highly specific IgM antibody capture ELISA (MAC-ELISA), which utilizes biotinylated recombinant SARS-CoV-2 nucleocapsid (N) antigen produced in plants. Our biotinylated approach streamlines the procedure by eliminating the requirement for an anti-N-conjugated antibody, circumventing the need for peroxidase-labeled antigens, and preventing cross-reactivity with IgM autoantibodies such as rheumatoid factor. Performance evaluation of the assay involved assessing sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy using 682 RT-qPCR-positive samples, categorized by weeks relative to symptoms onset. Negative controls included 205 pre-pandemic serum samples and 46 serum samples from patients diagnosed with other diseases. Based on a cut-off of 0.087 and ROC curve analysis, the highest sensitivity of 81.2% was observed in the 8-14 days post-symptom (dps) group (2nd week), followed by sensitivities of 73.8% and 68.37% for the 1-7 dps (1st week) and 15-21 dps groups (3rd week), respectively. Specificity was consistently 100% across all groups. This newly developed biotinylated N-MAC-ELISA offers a more streamlined and cost-effective alternative to molecular diagnostics. It enables simultaneous testing of multiple samples and effectively identifies individuals with false-negative results.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:54 |
|---|---|
| Enthalten in: |
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology - 54(2023), 4 vom: 06. Dez., Seite 2893-2901 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Lopes-Luz, Leonardo [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Antibodies, Viral |
|---|
| Anmerkungen: |
Date Completed 04.12.2023 Date Revised 05.12.2023 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1007/s42770-023-01160-6 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM364224479 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM364224479 | ||
| 003 | DE-627 | ||
| 005 | 20231226095008.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1007/s42770-023-01160-6 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1214.xml |
| 035 | |a (DE-627)NLM364224479 | ||
| 035 | |a (NLM)37930615 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Lopes-Luz, Leonardo |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A novel highly specific biotinylated MAC-ELISA for detection of anti-SARS-CoV-2 nucleocapsid antigen IgM antibodies during the acute phase of COVID-19 |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 04.12.2023 | ||
| 500 | |a Date Revised 05.12.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia. | ||
| 520 | |a The gold standard for diagnosing COVID-19 in the acute phase is RT-qPCR. However, this molecular technique can yield false-negative results when nasopharyngeal swab collection is not conducted during viremia. To mitigate this challenge, the enzyme-linked immunosorbent assay (ELISA) identifies anti-SARS-CoV-2 IgM antibodies in the initial weeks after symptom onset, facilitating early COVID-19 diagnosis. This study introduces a novel and highly specific IgM antibody capture ELISA (MAC-ELISA), which utilizes biotinylated recombinant SARS-CoV-2 nucleocapsid (N) antigen produced in plants. Our biotinylated approach streamlines the procedure by eliminating the requirement for an anti-N-conjugated antibody, circumventing the need for peroxidase-labeled antigens, and preventing cross-reactivity with IgM autoantibodies such as rheumatoid factor. Performance evaluation of the assay involved assessing sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy using 682 RT-qPCR-positive samples, categorized by weeks relative to symptoms onset. Negative controls included 205 pre-pandemic serum samples and 46 serum samples from patients diagnosed with other diseases. Based on a cut-off of 0.087 and ROC curve analysis, the highest sensitivity of 81.2% was observed in the 8-14 days post-symptom (dps) group (2nd week), followed by sensitivities of 73.8% and 68.37% for the 1-7 dps (1st week) and 15-21 dps groups (3rd week), respectively. Specificity was consistently 100% across all groups. This newly developed biotinylated N-MAC-ELISA offers a more streamlined and cost-effective alternative to molecular diagnostics. It enables simultaneous testing of multiple samples and effectively identifies individuals with false-negative results | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Biotinylated antigen | |
| 650 | 4 | |a IgM | |
| 650 | 4 | |a MAC-ELISA | |
| 650 | 4 | |a Rheumatoid factor | |
| 650 | 4 | |a Streptavidin | |
| 650 | 7 | |a Immunoglobulin M |2 NLM | |
| 650 | 7 | |a Antibodies, Viral |2 NLM | |
| 700 | 1 | |a Fogaça, Matheus Bernardes Torres |e verfasserin |4 aut | |
| 700 | 1 | |a Bentivoglio-Silva, Brenda Garcia |e verfasserin |4 aut | |
| 700 | 1 | |a Saavedra, Djairo Pastor |e verfasserin |4 aut | |
| 700 | 1 | |a Alves, Luana Michele |e verfasserin |4 aut | |
| 700 | 1 | |a Franca, Luísa Valério |e verfasserin |4 aut | |
| 700 | 1 | |a Crispim, Gildemar José Bezerra |e verfasserin |4 aut | |
| 700 | 1 | |a de Andrade, Ikaro Alves |e verfasserin |4 aut | |
| 700 | 1 | |a Ribeiro, Bergmann Morais |e verfasserin |4 aut | |
| 700 | 1 | |a Nagata, Tatsuya |e verfasserin |4 aut | |
| 700 | 1 | |a Bührer-Sékula, Samira |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology |d 2006 |g 54(2023), 4 vom: 06. Dez., Seite 2893-2901 |w (DE-627)NLM197858600 |x 1678-4405 |7 nnns |
| 773 | 1 | 8 | |g volume:54 |g year:2023 |g number:4 |g day:06 |g month:12 |g pages:2893-2901 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1007/s42770-023-01160-6 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 54 |j 2023 |e 4 |b 06 |c 12 |h 2893-2901 | ||