Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2
Polymerase chain reaction (PCR)-based antigen tests are technically difficult, time-consuming, and expensive, and may produce false negative results requiring follow-up confirmation with computed tomography. The global coronavirus disease 2019 (COVID-19) pandemic has increased the demand for accurat...
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doaj-fd29a4453a764d929ebde7a986a7c2b42020-11-25T03:35:04ZengMDPI AGDiagnostics2075-44182020-08-011059459410.3390/diagnostics10080594Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2Yuta Kyosei0Mayuri Namba1Sou Yamura2Rikiya Takeuchi3Noriko Aoki4Kazunari Nakaishi5Satoshi Watabe6Etsuro Ito7Department of Biology, Waseda University, 2-2 Wakamatsucho, Shinjuku, Tokyo 162-8480, JapanDepartment of Biology, Waseda University, 2-2 Wakamatsucho, Shinjuku, Tokyo 162-8480, JapanDepartment of Biology, Waseda University, 2-2 Wakamatsucho, Shinjuku, Tokyo 162-8480, JapanResearch and Development Department, TAUNS Laboratories, Inc., 245-1 Doniwa, Shimizu, Sunto, Shizuoka 411-0903, JapanResearch and Development Department, TAUNS Laboratories, Inc., 245-1 Doniwa, Shimizu, Sunto, Shizuoka 411-0903, JapanQuality Headquarters, TAUNS Laboratories, Inc., 761-1 Kamishima, Izunokuni, Shizuoka 410-2325, JapanResearch and Development Department, TAUNS Laboratories, Inc., 245-1 Doniwa, Shimizu, Sunto, Shizuoka 411-0903, JapanDepartment of Biology, Waseda University, 2-2 Wakamatsucho, Shinjuku, Tokyo 162-8480, JapanPolymerase chain reaction (PCR)-based antigen tests are technically difficult, time-consuming, and expensive, and may produce false negative results requiring follow-up confirmation with computed tomography. The global coronavirus disease 2019 (COVID-19) pandemic has increased the demand for accurate, easy-to-use, rapid, and cost-effective antigen tests for clinical application. We propose a de novo antigen test for diagnosing COVID-19 using the combination of sandwich enzyme-linked immunosorbent assay and thio-nicotinamide adenine dinucleotide (thio-NAD) cycling. Our test takes advantage of the spike proteins specific to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The limit of detection of our test was 2.3 × 10<sup>−18</sup> moles/assay. If the virus has ~25 spike proteins on its surface, our method should detect on the order of 10<sup>−20</sup> moles of virus/assay, corresponding to ~10<sup>4</sup> copies of the virus RNA/assay. The detection sensitivity approaches that of PCR-based assays because the average virus RNA load used for PCR-based assays is ~10<sup>5</sup> copies per oro- or naso-pharyngeal swab specimen. To our knowledge, this is the first ultrasensitive antigen test for SARS-CoV-2 spike proteins that can be performed with an easy-to-use microplate reader. Sufficient sensitivity can be achieved within 10 min of thio-NAD cycling. Our antigen test allows for rapid, cost-effective, specific, ultrasensitive, and simultaneous multiple measurements of SARS-CoV-2, and has broad application for the diagnosis for COVID-19.https://www.mdpi.com/2075-4418/10/8/594COVID-19SARS-CoV-2spike proteinthio-NAD cyclingultrasensitive ELISA |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuta Kyosei Mayuri Namba Sou Yamura Rikiya Takeuchi Noriko Aoki Kazunari Nakaishi Satoshi Watabe Etsuro Ito |
spellingShingle |
Yuta Kyosei Mayuri Namba Sou Yamura Rikiya Takeuchi Noriko Aoki Kazunari Nakaishi Satoshi Watabe Etsuro Ito Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2 Diagnostics COVID-19 SARS-CoV-2 spike protein thio-NAD cycling ultrasensitive ELISA |
author_facet |
Yuta Kyosei Mayuri Namba Sou Yamura Rikiya Takeuchi Noriko Aoki Kazunari Nakaishi Satoshi Watabe Etsuro Ito |
author_sort |
Yuta Kyosei |
title |
Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2 |
title_short |
Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2 |
title_full |
Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2 |
title_fullStr |
Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2 |
title_full_unstemmed |
Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2 |
title_sort |
proposal of de novo antigen test for covid-19: ultrasensitive detection of spike proteins of sars-cov-2 |
publisher |
MDPI AG |
series |
Diagnostics |
issn |
2075-4418 |
publishDate |
2020-08-01 |
description |
Polymerase chain reaction (PCR)-based antigen tests are technically difficult, time-consuming, and expensive, and may produce false negative results requiring follow-up confirmation with computed tomography. The global coronavirus disease 2019 (COVID-19) pandemic has increased the demand for accurate, easy-to-use, rapid, and cost-effective antigen tests for clinical application. We propose a de novo antigen test for diagnosing COVID-19 using the combination of sandwich enzyme-linked immunosorbent assay and thio-nicotinamide adenine dinucleotide (thio-NAD) cycling. Our test takes advantage of the spike proteins specific to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The limit of detection of our test was 2.3 × 10<sup>−18</sup> moles/assay. If the virus has ~25 spike proteins on its surface, our method should detect on the order of 10<sup>−20</sup> moles of virus/assay, corresponding to ~10<sup>4</sup> copies of the virus RNA/assay. The detection sensitivity approaches that of PCR-based assays because the average virus RNA load used for PCR-based assays is ~10<sup>5</sup> copies per oro- or naso-pharyngeal swab specimen. To our knowledge, this is the first ultrasensitive antigen test for SARS-CoV-2 spike proteins that can be performed with an easy-to-use microplate reader. Sufficient sensitivity can be achieved within 10 min of thio-NAD cycling. Our antigen test allows for rapid, cost-effective, specific, ultrasensitive, and simultaneous multiple measurements of SARS-CoV-2, and has broad application for the diagnosis for COVID-19. |
topic |
COVID-19 SARS-CoV-2 spike protein thio-NAD cycling ultrasensitive ELISA |
url |
https://www.mdpi.com/2075-4418/10/8/594 |
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