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Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2
The severe acute respiratory syndrome coronavirus (SARS-CoV-2) has infected millions of individuals and continues to be a major health concern worldwide. While reverse transcription-polymerase chain reaction remains a reliable method for detecting infections, limitations of this technology, particul...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Author(s). Published by Elsevier B.V.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10223630/ https://www.ncbi.nlm.nih.gov/pubmed/37275459 http://dx.doi.org/10.1016/j.crbiot.2023.100132 |
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author | Neff, Charles P. Cikara, Mile Geiss, Brian J. Thomas Caltagirone, G. Liao, Albert Atif, Shaikh M. Macdonald, Bradley Schaden, Richard |
author_facet | Neff, Charles P. Cikara, Mile Geiss, Brian J. Thomas Caltagirone, G. Liao, Albert Atif, Shaikh M. Macdonald, Bradley Schaden, Richard |
author_sort | Neff, Charles P. |
collection | PubMed |
description | The severe acute respiratory syndrome coronavirus (SARS-CoV-2) has infected millions of individuals and continues to be a major health concern worldwide. While reverse transcription-polymerase chain reaction remains a reliable method for detecting infections, limitations of this technology, particularly cost and the requirement of a dedicated laboratory, prevent rapid viral monitoring. Antigen tests filled this need to some extent but with limitations including sensitivity and specificity, particularly against emerging variants of concern. Here, we developed aptamers against the SARS-CoV-2 Nucleocapsid protein to complement or replace antibodies in antigen detection assays. As detection reagents in ELISA-like assays, our DNA aptamers were able to detect as low as 150 pg/mL of the protein and under 150 k copies of inactivated SARS-CoV-2 Wuhan Alpha strain in viral transport medium with little cross-reactivity to other human coronaviruses (HCoVs). Further, our aptamers were reselected against the SARS-CoV-2 Omicron variant of concern, and we found two sequences that had a more than two-fold increase in signal compared to our original aptamers when used as detection reagents against protein from the Omicron strain. These findings illustrate the use of aptamers as promising alternative detection reagents that may translate for use in current tests and our findings validate the method for the reselection of aptamers against emerging viral strains. |
format | Online Article Text |
id | pubmed-10223630 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The Author(s). Published by Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-102236302023-05-30 Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 Neff, Charles P. Cikara, Mile Geiss, Brian J. Thomas Caltagirone, G. Liao, Albert Atif, Shaikh M. Macdonald, Bradley Schaden, Richard Curr Res Biotechnol Article The severe acute respiratory syndrome coronavirus (SARS-CoV-2) has infected millions of individuals and continues to be a major health concern worldwide. While reverse transcription-polymerase chain reaction remains a reliable method for detecting infections, limitations of this technology, particularly cost and the requirement of a dedicated laboratory, prevent rapid viral monitoring. Antigen tests filled this need to some extent but with limitations including sensitivity and specificity, particularly against emerging variants of concern. Here, we developed aptamers against the SARS-CoV-2 Nucleocapsid protein to complement or replace antibodies in antigen detection assays. As detection reagents in ELISA-like assays, our DNA aptamers were able to detect as low as 150 pg/mL of the protein and under 150 k copies of inactivated SARS-CoV-2 Wuhan Alpha strain in viral transport medium with little cross-reactivity to other human coronaviruses (HCoVs). Further, our aptamers were reselected against the SARS-CoV-2 Omicron variant of concern, and we found two sequences that had a more than two-fold increase in signal compared to our original aptamers when used as detection reagents against protein from the Omicron strain. These findings illustrate the use of aptamers as promising alternative detection reagents that may translate for use in current tests and our findings validate the method for the reselection of aptamers against emerging viral strains. The Author(s). Published by Elsevier B.V. 2023 2023-05-27 /pmc/articles/PMC10223630/ /pubmed/37275459 http://dx.doi.org/10.1016/j.crbiot.2023.100132 Text en © 2023 The Author(s) Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Neff, Charles P. Cikara, Mile Geiss, Brian J. Thomas Caltagirone, G. Liao, Albert Atif, Shaikh M. Macdonald, Bradley Schaden, Richard Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 |
title | Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 |
title_full | Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 |
title_fullStr | Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 |
title_full_unstemmed | Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 |
title_short | Nucleocapsid protein binding DNA aptamers for detection of SARS-COV-2 |
title_sort | nucleocapsid protein binding dna aptamers for detection of sars-cov-2 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10223630/ https://www.ncbi.nlm.nih.gov/pubmed/37275459 http://dx.doi.org/10.1016/j.crbiot.2023.100132 |
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