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Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection

The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 with...

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Detalles Bibliográficos
Autores principales: Barza, Ruby, Patel, Parul, Sabatini, Linda, Singh, Kamaljit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7418644/
https://www.ncbi.nlm.nih.gov/pubmed/32898817
http://dx.doi.org/10.1016/j.jcv.2020.104587
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author Barza, Ruby
Patel, Parul
Sabatini, Linda
Singh, Kamaljit
author_facet Barza, Ruby
Patel, Parul
Sabatini, Linda
Singh, Kamaljit
author_sort Barza, Ruby
collection PubMed
description The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 without the need for nucleic acid extraction. RT-PCR results using the ChromaCode HDPCR™ SARS-CoV-2 were compared using the heat-RNA release method and an automated RNA extraction system (EMAG). The heat-RNA release method correctly identified 94 % (81/86 nasopharyngeal samples) that were positive for SARS-CoV-2. Five samples that were missed by heat-RNA release method had a mean Ct value: 35 using the automated extraction instrument, indicating a very low viral load. Our findings show that a simple heat-RNA release method is a reasonable alternative for the majority of COVID-19 positive patients and can help overcome the cost and availability issues of RNA extraction reagents.
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spelling pubmed-74186442020-08-12 Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection Barza, Ruby Patel, Parul Sabatini, Linda Singh, Kamaljit J Clin Virol Short Communication The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 without the need for nucleic acid extraction. RT-PCR results using the ChromaCode HDPCR™ SARS-CoV-2 were compared using the heat-RNA release method and an automated RNA extraction system (EMAG). The heat-RNA release method correctly identified 94 % (81/86 nasopharyngeal samples) that were positive for SARS-CoV-2. Five samples that were missed by heat-RNA release method had a mean Ct value: 35 using the automated extraction instrument, indicating a very low viral load. Our findings show that a simple heat-RNA release method is a reasonable alternative for the majority of COVID-19 positive patients and can help overcome the cost and availability issues of RNA extraction reagents. Elsevier B.V. 2020-11 2020-08-11 /pmc/articles/PMC7418644/ /pubmed/32898817 http://dx.doi.org/10.1016/j.jcv.2020.104587 Text en © 2020 Elsevier B.V. All rights reserved. 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 Short Communication
Barza, Ruby
Patel, Parul
Sabatini, Linda
Singh, Kamaljit
Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
title Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
title_full Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
title_fullStr Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
title_full_unstemmed Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
title_short Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
title_sort use of a simplified sample processing step without rna extraction for direct sars-cov-2 rt-pcr detection
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7418644/
https://www.ncbi.nlm.nih.gov/pubmed/32898817
http://dx.doi.org/10.1016/j.jcv.2020.104587
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