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SARS-CoV-2: Comparative analysis of different RNA extraction methods
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of the COVID-19 pandemic. Although other diagnostic methods have been introduced, detection of viral genes on oro- and nasopharyngeal swabs by reverse-transcription real time-PCR (rRT-PCR) assays is still the gold st...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Author(s). Published by Elsevier B.V.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7640895/ https://www.ncbi.nlm.nih.gov/pubmed/33160015 http://dx.doi.org/10.1016/j.jviromet.2020.114008 |
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author | Ambrosi, Cecilia Prezioso, Carla Checconi, Paola Scribano, Daniela Sarshar, Meysam Capannari, Maurizio Tomino, Carlo Fini, Massimo Garaci, Enrico Palamara, Anna Teresa De Chiara, Giovanna Limongi, Dolores |
author_facet | Ambrosi, Cecilia Prezioso, Carla Checconi, Paola Scribano, Daniela Sarshar, Meysam Capannari, Maurizio Tomino, Carlo Fini, Massimo Garaci, Enrico Palamara, Anna Teresa De Chiara, Giovanna Limongi, Dolores |
author_sort | Ambrosi, Cecilia |
collection | PubMed |
description | Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of the COVID-19 pandemic. Although other diagnostic methods have been introduced, detection of viral genes on oro- and nasopharyngeal swabs by reverse-transcription real time-PCR (rRT-PCR) assays is still the gold standard. Efficient viral RNA extraction is a prerequisite for downstream performance of rRT-PCR assays. Currently, several automatic methods that include RNA extraction are available. However, due to the growing demand, a shortage in kit supplies could be experienced in several labs. For these reasons, the use of different commercial or in-house protocols for RNA extraction may increase the possibility to analyze high number of samples. Herein, we compared the efficiency of RNA extraction of three different commercial kits and an in-house extraction protocol using synthetic ssRNA standards of SARS-CoV-2 as well as in oro-nasopharyngeal swabs from six COVID-19-positive patients. It was concluded that tested commercial kits can be used with some modifications for the detection of the SARS-CoV-2 genome by rRT-PCR approaches, although with some differences in RNA yields. Conversely, EXTRAzol reagent was the less efficient due to the phase separation principle at the basis of RNA extraction. Overall, this study offers alternative suitable methods to manually extract RNA that can be taken into account for SARS-CoV-2 detection. |
format | Online Article Text |
id | pubmed-7640895 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Author(s). Published by Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-76408952020-11-05 SARS-CoV-2: Comparative analysis of different RNA extraction methods Ambrosi, Cecilia Prezioso, Carla Checconi, Paola Scribano, Daniela Sarshar, Meysam Capannari, Maurizio Tomino, Carlo Fini, Massimo Garaci, Enrico Palamara, Anna Teresa De Chiara, Giovanna Limongi, Dolores J Virol Methods Article Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of the COVID-19 pandemic. Although other diagnostic methods have been introduced, detection of viral genes on oro- and nasopharyngeal swabs by reverse-transcription real time-PCR (rRT-PCR) assays is still the gold standard. Efficient viral RNA extraction is a prerequisite for downstream performance of rRT-PCR assays. Currently, several automatic methods that include RNA extraction are available. However, due to the growing demand, a shortage in kit supplies could be experienced in several labs. For these reasons, the use of different commercial or in-house protocols for RNA extraction may increase the possibility to analyze high number of samples. Herein, we compared the efficiency of RNA extraction of three different commercial kits and an in-house extraction protocol using synthetic ssRNA standards of SARS-CoV-2 as well as in oro-nasopharyngeal swabs from six COVID-19-positive patients. It was concluded that tested commercial kits can be used with some modifications for the detection of the SARS-CoV-2 genome by rRT-PCR approaches, although with some differences in RNA yields. Conversely, EXTRAzol reagent was the less efficient due to the phase separation principle at the basis of RNA extraction. Overall, this study offers alternative suitable methods to manually extract RNA that can be taken into account for SARS-CoV-2 detection. The Author(s). Published by Elsevier B.V. 2021-01 2020-11-04 /pmc/articles/PMC7640895/ /pubmed/33160015 http://dx.doi.org/10.1016/j.jviromet.2020.114008 Text en © 2020 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 Ambrosi, Cecilia Prezioso, Carla Checconi, Paola Scribano, Daniela Sarshar, Meysam Capannari, Maurizio Tomino, Carlo Fini, Massimo Garaci, Enrico Palamara, Anna Teresa De Chiara, Giovanna Limongi, Dolores SARS-CoV-2: Comparative analysis of different RNA extraction methods |
title | SARS-CoV-2: Comparative analysis of different RNA extraction methods |
title_full | SARS-CoV-2: Comparative analysis of different RNA extraction methods |
title_fullStr | SARS-CoV-2: Comparative analysis of different RNA extraction methods |
title_full_unstemmed | SARS-CoV-2: Comparative analysis of different RNA extraction methods |
title_short | SARS-CoV-2: Comparative analysis of different RNA extraction methods |
title_sort | sars-cov-2: comparative analysis of different rna extraction methods |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7640895/ https://www.ncbi.nlm.nih.gov/pubmed/33160015 http://dx.doi.org/10.1016/j.jviromet.2020.114008 |
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