Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials

To help control the global pandemic of coronavirus disease 2019 (COVID-19), we developed a diagnostic method targeting the spike protein of the virus that causes the infection, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We applied an ultrasensitive method by combining a sandwich e...

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Autores principales:	Yuta Kyosei, Mayuri Namba, Daiki Makioka, Ayumi Kokubun, Satoshi Watabe, Teruki Yoshimura, Tadahiro Sasaki, Tatsuo Shioda, Etsuro Ito
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	antigen test COVID-19 SARS-CoV-2 spike protein thio-NAD cycling ultrasensitive ELISA Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/1ace5b5ef65c484787afd1376fd416ba
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spelling	oai:doaj.org-article:1ace5b5ef65c484787afd1376fd416ba2021-11-25T18:24:18ZUltrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials10.3390/microorganisms91122142076-2607https://doaj.org/article/1ace5b5ef65c484787afd1376fd416ba2021-10-01T00:00:00Zhttps://www.mdpi.com/2076-2607/9/11/2214https://doaj.org/toc/2076-2607To help control the global pandemic of coronavirus disease 2019 (COVID-19), we developed a diagnostic method targeting the spike protein of the virus that causes the infection, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We applied an ultrasensitive method by combining a sandwich enzyme-linked immunosorbent assay (ELISA) and the thio-nicotinamide adenine dinucleotide (thio-NAD) cycling reaction to quantify spike S1 proteins. The limit of detection (LOD) was 2.62 × 10<sup>−19</sup> moles/assay for recombinant S1 proteins and 2.6 × 10<sup>6</sup> RNA copies/assay for ultraviolet B-inactivated viruses. We have already shown that the ultrasensitive ELISA for nucleocapsid proteins can detect ultraviolet B-inactivated viruses at the 10<sup>4</sup> RNA copies/assay level, whereas the nucleocapsid proteins of SARS-CoV-2 are difficult to distinguish from those in conventional coronaviruses and SARS-CoV. Thus, an antigen test for only the nucleocapsid proteins is insufficient for virus specificity. Therefore, the use of a combination of tests against both spike and nucleocapsid proteins is recommended to increase both the detection sensitivity and testing accuracy of the COVID-19 antigen test. Taken together, our present study, in which we incorporate S1 detection by combining the ultrasensitive ELISA for nucleocapsid proteins, offers an ultrasensitive, antigen-specific test for COVID-19.Yuta KyoseiMayuri NambaDaiki MakiokaAyumi KokubunSatoshi WatabeTeruki YoshimuraTadahiro SasakiTatsuo ShiodaEtsuro ItoMDPI AGarticleantigen testCOVID-19SARS-CoV-2spike proteinthio-NAD cyclingultrasensitive ELISABiology (General)QH301-705.5ENMicroorganisms, Vol 9, Iss 2214, p 2214 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	antigen test COVID-19 SARS-CoV-2 spike protein thio-NAD cycling ultrasensitive ELISA Biology (General) QH301-705.5
spellingShingle	antigen test COVID-19 SARS-CoV-2 spike protein thio-NAD cycling ultrasensitive ELISA Biology (General) QH301-705.5 Yuta Kyosei Mayuri Namba Daiki Makioka Ayumi Kokubun Satoshi Watabe Teruki Yoshimura Tadahiro Sasaki Tatsuo Shioda Etsuro Ito Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials
description	To help control the global pandemic of coronavirus disease 2019 (COVID-19), we developed a diagnostic method targeting the spike protein of the virus that causes the infection, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We applied an ultrasensitive method by combining a sandwich enzyme-linked immunosorbent assay (ELISA) and the thio-nicotinamide adenine dinucleotide (thio-NAD) cycling reaction to quantify spike S1 proteins. The limit of detection (LOD) was 2.62 × 10<sup>−19</sup> moles/assay for recombinant S1 proteins and 2.6 × 10<sup>6</sup> RNA copies/assay for ultraviolet B-inactivated viruses. We have already shown that the ultrasensitive ELISA for nucleocapsid proteins can detect ultraviolet B-inactivated viruses at the 10<sup>4</sup> RNA copies/assay level, whereas the nucleocapsid proteins of SARS-CoV-2 are difficult to distinguish from those in conventional coronaviruses and SARS-CoV. Thus, an antigen test for only the nucleocapsid proteins is insufficient for virus specificity. Therefore, the use of a combination of tests against both spike and nucleocapsid proteins is recommended to increase both the detection sensitivity and testing accuracy of the COVID-19 antigen test. Taken together, our present study, in which we incorporate S1 detection by combining the ultrasensitive ELISA for nucleocapsid proteins, offers an ultrasensitive, antigen-specific test for COVID-19.
format	article
author	Yuta Kyosei Mayuri Namba Daiki Makioka Ayumi Kokubun Satoshi Watabe Teruki Yoshimura Tadahiro Sasaki Tatsuo Shioda Etsuro Ito
author_facet	Yuta Kyosei Mayuri Namba Daiki Makioka Ayumi Kokubun Satoshi Watabe Teruki Yoshimura Tadahiro Sasaki Tatsuo Shioda Etsuro Ito
author_sort	Yuta Kyosei
title	Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials
title_short	Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials
title_full	Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials
title_fullStr	Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials
title_full_unstemmed	Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials
title_sort	ultrasensitive detection of sars-cov-2 spike proteins using the thio-nad cycling reaction: a preliminary study before clinical trials
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/1ace5b5ef65c484787afd1376fd416ba
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Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials

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