Cargando…

Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants

The COVID-19 pandemic highlights the need for diagnostics that can be rapidly adapted and deployed in a variety of settings. Several SARS-CoV-2 variants have shown worrisome effects on vaccine and treatment efficacy, but no current point-of-care (POC) testing modality allows their specific identific...

Descripción completa

Detalles Bibliográficos
Autores principales: de Puig, Helena, Lee, Rose A., Najjar, Devora, Tan, Xiao, Soekensen, Luis R., Angenent-Mari, Nicolaas M., Donghia, Nina M., Weckman, Nicole E., Ory, Audrey, Ng, Carlos F., Nguyen, Peter Q., Mao, Angelo S., Ferrante, Thomas C., Lansberry, Geoffrey, Sallum, Hani, Niemi, James, Collins, James J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8346217/
https://www.ncbi.nlm.nih.gov/pubmed/34362739
http://dx.doi.org/10.1126/sciadv.abh2944
_version_ 1783734818656223232
author de Puig, Helena
Lee, Rose A.
Najjar, Devora
Tan, Xiao
Soekensen, Luis R.
Angenent-Mari, Nicolaas M.
Donghia, Nina M.
Weckman, Nicole E.
Ory, Audrey
Ng, Carlos F.
Nguyen, Peter Q.
Mao, Angelo S.
Ferrante, Thomas C.
Lansberry, Geoffrey
Sallum, Hani
Niemi, James
Collins, James J.
author_facet de Puig, Helena
Lee, Rose A.
Najjar, Devora
Tan, Xiao
Soekensen, Luis R.
Angenent-Mari, Nicolaas M.
Donghia, Nina M.
Weckman, Nicole E.
Ory, Audrey
Ng, Carlos F.
Nguyen, Peter Q.
Mao, Angelo S.
Ferrante, Thomas C.
Lansberry, Geoffrey
Sallum, Hani
Niemi, James
Collins, James J.
author_sort de Puig, Helena
collection PubMed
description The COVID-19 pandemic highlights the need for diagnostics that can be rapidly adapted and deployed in a variety of settings. Several SARS-CoV-2 variants have shown worrisome effects on vaccine and treatment efficacy, but no current point-of-care (POC) testing modality allows their specific identification. We have developed miSHERLOCK, a low-cost, CRISPR-based POC diagnostic platform that takes unprocessed patient saliva; extracts, purifies, and concentrates viral RNA; performs amplification and detection reactions; and provides fluorescent visual output with only three user actions and 1 hour from sample input to answer out. miSHERLOCK achieves highly sensitive multiplexed detection of SARS-CoV-2 and mutations associated with variants B.1.1.7, B.1.351, and P.1. Our modular system enables easy exchange of assays to address diverse user needs and can be rapidly reconfigured to detect different viruses and variants of concern. An adjunctive smartphone application enables output quantification, automated interpretation, and the possibility of remote, distributed result reporting.
format Online
Article
Text
id pubmed-8346217
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher American Association for the Advancement of Science
record_format MEDLINE/PubMed
spelling pubmed-83462172021-08-13 Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants de Puig, Helena Lee, Rose A. Najjar, Devora Tan, Xiao Soekensen, Luis R. Angenent-Mari, Nicolaas M. Donghia, Nina M. Weckman, Nicole E. Ory, Audrey Ng, Carlos F. Nguyen, Peter Q. Mao, Angelo S. Ferrante, Thomas C. Lansberry, Geoffrey Sallum, Hani Niemi, James Collins, James J. Sci Adv Research Articles The COVID-19 pandemic highlights the need for diagnostics that can be rapidly adapted and deployed in a variety of settings. Several SARS-CoV-2 variants have shown worrisome effects on vaccine and treatment efficacy, but no current point-of-care (POC) testing modality allows their specific identification. We have developed miSHERLOCK, a low-cost, CRISPR-based POC diagnostic platform that takes unprocessed patient saliva; extracts, purifies, and concentrates viral RNA; performs amplification and detection reactions; and provides fluorescent visual output with only three user actions and 1 hour from sample input to answer out. miSHERLOCK achieves highly sensitive multiplexed detection of SARS-CoV-2 and mutations associated with variants B.1.1.7, B.1.351, and P.1. Our modular system enables easy exchange of assays to address diverse user needs and can be rapidly reconfigured to detect different viruses and variants of concern. An adjunctive smartphone application enables output quantification, automated interpretation, and the possibility of remote, distributed result reporting. American Association for the Advancement of Science 2021-08-06 /pmc/articles/PMC8346217/ /pubmed/34362739 http://dx.doi.org/10.1126/sciadv.abh2944 Text en Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). https://creativecommons.org/licenses/by-nc/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (https://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.
spellingShingle Research Articles
de Puig, Helena
Lee, Rose A.
Najjar, Devora
Tan, Xiao
Soekensen, Luis R.
Angenent-Mari, Nicolaas M.
Donghia, Nina M.
Weckman, Nicole E.
Ory, Audrey
Ng, Carlos F.
Nguyen, Peter Q.
Mao, Angelo S.
Ferrante, Thomas C.
Lansberry, Geoffrey
Sallum, Hani
Niemi, James
Collins, James J.
Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants
title Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants
title_full Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants
title_fullStr Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants
title_full_unstemmed Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants
title_short Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants
title_sort minimally instrumented sherlock (misherlock) for crispr-based point-of-care diagnosis of sars-cov-2 and emerging variants
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8346217/
https://www.ncbi.nlm.nih.gov/pubmed/34362739
http://dx.doi.org/10.1126/sciadv.abh2944
work_keys_str_mv AT depuighelena minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT leerosea minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT najjardevora minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT tanxiao minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT soekensenluisr minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT angenentmarinicolaasm minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT donghianinam minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT weckmannicolee minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT oryaudrey minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT ngcarlosf minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT nguyenpeterq minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT maoangelos minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT ferrantethomasc minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT lansberrygeoffrey minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT sallumhani minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT niemijames minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants
AT collinsjamesj minimallyinstrumentedsherlockmisherlockforcrisprbasedpointofcarediagnosisofsarscov2andemergingvariants