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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...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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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 |
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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 |
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