Cargando…

A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples

Molecular diagnosis and surveillance of pathogens such as SARS‐CoV‐2 depend on nucleic acid isolation. Pandemics at the scale of COVID‐19 can cause a global shortage of proprietary commercial reagents and BSL‐2 laboratories to safely perform testing. Therefore, alternative solutions are urgently nee...

Descripción completa

Detalles Bibliográficos
Autores principales: Ramos‐Mandujano, Gerardo, Salunke, Rahul, Mfarrej, Sara, Rachmadi, Andri Taruna, Hala, Sharif, Xu, Jinna, Alofi, Fadwa S., Khogeer, Asim, Hashem, Anwar M., Almontashiri, Naif A. M., Alsomali, Afrah, Shinde, Digambar B., Hamdan, Samir, Hong, Pei‐Ying, Pain, Arnab, Li, Mo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7995109/
https://www.ncbi.nlm.nih.gov/pubmed/33786197
http://dx.doi.org/10.1002/gch2.202000068
_version_ 1783669875146752000
author Ramos‐Mandujano, Gerardo
Salunke, Rahul
Mfarrej, Sara
Rachmadi, Andri Taruna
Hala, Sharif
Xu, Jinna
Alofi, Fadwa S.
Khogeer, Asim
Hashem, Anwar M.
Almontashiri, Naif A. M.
Alsomali, Afrah
Shinde, Digambar B.
Hamdan, Samir
Hong, Pei‐Ying
Pain, Arnab
Li, Mo
author_facet Ramos‐Mandujano, Gerardo
Salunke, Rahul
Mfarrej, Sara
Rachmadi, Andri Taruna
Hala, Sharif
Xu, Jinna
Alofi, Fadwa S.
Khogeer, Asim
Hashem, Anwar M.
Almontashiri, Naif A. M.
Alsomali, Afrah
Shinde, Digambar B.
Hamdan, Samir
Hong, Pei‐Ying
Pain, Arnab
Li, Mo
author_sort Ramos‐Mandujano, Gerardo
collection PubMed
description Molecular diagnosis and surveillance of pathogens such as SARS‐CoV‐2 depend on nucleic acid isolation. Pandemics at the scale of COVID‐19 can cause a global shortage of proprietary commercial reagents and BSL‐2 laboratories to safely perform testing. Therefore, alternative solutions are urgently needed to address these challenges. An open‐source method, magnetic‐nanoparticle‐aided viral RNA isolation from contagious samples (MAVRICS), built upon readily available reagents, and easily assembled in any basically equipped laboratory, is thus developed. The performance of MAVRICS is evaluated using validated pathogen detection assays and real‐world and contrived samples. Unlike conventional methods, MAVRICS works directly in samples inactivated in phenol‐chloroform (e.g., TRIzol), thus allowing infectious samples to be handled safely without biocontainment facilities. MAVRICS allows wastewater biomass immobilized on membranes to be directly inactivated and lysed in TRIzol followed by RNA extraction by magnetic nanoparticles, thereby greatly reducing biohazard risk and simplifying processing procedures. Using 39 COVID‐19 patient samples and two wastewater samples, it is shown that MAVRICS rivals commercial kits in detection of SARS‐CoV‐2, influenza viruses, and respiratory syncytial virus. Therefore, MAVRICS is safe, fast, and scalable. It is field‐deployable with minimal equipment requirements and could become an enabling technology for widespread testing and wastewater monitoring of diverse pathogens.
format Online
Article
Text
id pubmed-7995109
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-79951092021-03-26 A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples Ramos‐Mandujano, Gerardo Salunke, Rahul Mfarrej, Sara Rachmadi, Andri Taruna Hala, Sharif Xu, Jinna Alofi, Fadwa S. Khogeer, Asim Hashem, Anwar M. Almontashiri, Naif A. M. Alsomali, Afrah Shinde, Digambar B. Hamdan, Samir Hong, Pei‐Ying Pain, Arnab Li, Mo Glob Chall Full Papers Molecular diagnosis and surveillance of pathogens such as SARS‐CoV‐2 depend on nucleic acid isolation. Pandemics at the scale of COVID‐19 can cause a global shortage of proprietary commercial reagents and BSL‐2 laboratories to safely perform testing. Therefore, alternative solutions are urgently needed to address these challenges. An open‐source method, magnetic‐nanoparticle‐aided viral RNA isolation from contagious samples (MAVRICS), built upon readily available reagents, and easily assembled in any basically equipped laboratory, is thus developed. The performance of MAVRICS is evaluated using validated pathogen detection assays and real‐world and contrived samples. Unlike conventional methods, MAVRICS works directly in samples inactivated in phenol‐chloroform (e.g., TRIzol), thus allowing infectious samples to be handled safely without biocontainment facilities. MAVRICS allows wastewater biomass immobilized on membranes to be directly inactivated and lysed in TRIzol followed by RNA extraction by magnetic nanoparticles, thereby greatly reducing biohazard risk and simplifying processing procedures. Using 39 COVID‐19 patient samples and two wastewater samples, it is shown that MAVRICS rivals commercial kits in detection of SARS‐CoV‐2, influenza viruses, and respiratory syncytial virus. Therefore, MAVRICS is safe, fast, and scalable. It is field‐deployable with minimal equipment requirements and could become an enabling technology for widespread testing and wastewater monitoring of diverse pathogens. John Wiley and Sons Inc. 2021-02-22 /pmc/articles/PMC7995109/ /pubmed/33786197 http://dx.doi.org/10.1002/gch2.202000068 Text en © 2021 The Authors. Global Challenges published by Wiley‐VCH GmbH This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Full Papers
Ramos‐Mandujano, Gerardo
Salunke, Rahul
Mfarrej, Sara
Rachmadi, Andri Taruna
Hala, Sharif
Xu, Jinna
Alofi, Fadwa S.
Khogeer, Asim
Hashem, Anwar M.
Almontashiri, Naif A. M.
Alsomali, Afrah
Shinde, Digambar B.
Hamdan, Samir
Hong, Pei‐Ying
Pain, Arnab
Li, Mo
A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples
title A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples
title_full A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples
title_fullStr A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples
title_full_unstemmed A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples
title_short A Robust, Safe, and Scalable Magnetic Nanoparticle Workflow for RNA Extraction of Pathogens from Clinical and Wastewater Samples
title_sort robust, safe, and scalable magnetic nanoparticle workflow for rna extraction of pathogens from clinical and wastewater samples
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7995109/
https://www.ncbi.nlm.nih.gov/pubmed/33786197
http://dx.doi.org/10.1002/gch2.202000068
work_keys_str_mv AT ramosmandujanogerardo arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT salunkerahul arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT mfarrejsara arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT rachmadiandritaruna arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT halasharif arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT xujinna arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT alofifadwas arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT khogeerasim arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT hashemanwarm arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT almontashirinaifam arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT alsomaliafrah arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT shindedigambarb arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT hamdansamir arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT hongpeiying arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT painarnab arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT limo arobustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT ramosmandujanogerardo robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT salunkerahul robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT mfarrejsara robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT rachmadiandritaruna robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT halasharif robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT xujinna robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT alofifadwas robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT khogeerasim robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT hashemanwarm robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT almontashirinaifam robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT alsomaliafrah robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT shindedigambarb robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT hamdansamir robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT hongpeiying robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT painarnab robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples
AT limo robustsafeandscalablemagneticnanoparticleworkflowforrnaextractionofpathogensfromclinicalandwastewatersamples