Cargando…

Comparison of DNA extraction methods for COVID-19 host genetics studies

The coronavirus disease 2019 (COVID-19) pandemic has resulted in global shortages in supplies for diagnostic tests, especially in the developing world. Risk factors for COVID-19 severity include pre-existing comorbidities, older age and male sex, but other variables are likely play a role in disease...

Descripción completa

Detalles Bibliográficos
Autores principales: da Silva, Ronaldo Celerino, de Lima, Suelen Cristina, dos Santos Reis, Wendell Palôma Maria, de Magalhães, Jurandy Júnior Ferraz, Magalhães, Ronaldo Nascimento de Oliveira, Rathi, Brijesh, Kohl, Alain, Bezerra, Marcos André Cavalcanti, Pena, Lindomar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10615309/
https://www.ncbi.nlm.nih.gov/pubmed/37903126
http://dx.doi.org/10.1371/journal.pone.0287551
_version_ 1785129194882072576
author da Silva, Ronaldo Celerino
de Lima, Suelen Cristina
dos Santos Reis, Wendell Palôma Maria
de Magalhães, Jurandy Júnior Ferraz
Magalhães, Ronaldo Nascimento de Oliveira
Rathi, Brijesh
Kohl, Alain
Bezerra, Marcos André Cavalcanti
Pena, Lindomar
author_facet da Silva, Ronaldo Celerino
de Lima, Suelen Cristina
dos Santos Reis, Wendell Palôma Maria
de Magalhães, Jurandy Júnior Ferraz
Magalhães, Ronaldo Nascimento de Oliveira
Rathi, Brijesh
Kohl, Alain
Bezerra, Marcos André Cavalcanti
Pena, Lindomar
author_sort da Silva, Ronaldo Celerino
collection PubMed
description The coronavirus disease 2019 (COVID-19) pandemic has resulted in global shortages in supplies for diagnostic tests, especially in the developing world. Risk factors for COVID-19 severity include pre-existing comorbidities, older age and male sex, but other variables are likely play a role in disease outcome. There is indeed increasing evidence that supports the role of host genetics in the predisposition to COVID-19 outcomes. The identification of genetic factors associated with the course of SARS-CoV-2 infections relies on DNA extraction methods. This study compared three DNA extraction methods (Chelex(®)100 resin, phenol-chloroform and the QIAamp DNA extraction kit) for COVID-19 host genetic studies using nasopharyngeal samples from patients. The methods were compared regarding number of required steps for execution, sample handling time, quality and quantity of the extracted material and application in genetic studies. The Chelex(®)100 method was found to be cheapest (33 and 13 times cheaper than the commercial kit and phenol-chloroform, respectively), give the highest DNA yield (306 and 69 times higher than the commercial kit and phenol-chloroform, respectively), with the least handling steps while providing adequate DNA quality for downstream applications. Together, our results show that the Chelex(®)100 resin is an inexpensive, safe, simple, fast, and suitable method for DNA extraction of nasopharyngeal samples from COVID-19 patients for genetics studies. This is particularly relevant in developing countries where cost and handling are critical steps in material processing.
format Online
Article
Text
id pubmed-10615309
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-106153092023-10-31 Comparison of DNA extraction methods for COVID-19 host genetics studies da Silva, Ronaldo Celerino de Lima, Suelen Cristina dos Santos Reis, Wendell Palôma Maria de Magalhães, Jurandy Júnior Ferraz Magalhães, Ronaldo Nascimento de Oliveira Rathi, Brijesh Kohl, Alain Bezerra, Marcos André Cavalcanti Pena, Lindomar PLoS One Research Article The coronavirus disease 2019 (COVID-19) pandemic has resulted in global shortages in supplies for diagnostic tests, especially in the developing world. Risk factors for COVID-19 severity include pre-existing comorbidities, older age and male sex, but other variables are likely play a role in disease outcome. There is indeed increasing evidence that supports the role of host genetics in the predisposition to COVID-19 outcomes. The identification of genetic factors associated with the course of SARS-CoV-2 infections relies on DNA extraction methods. This study compared three DNA extraction methods (Chelex(®)100 resin, phenol-chloroform and the QIAamp DNA extraction kit) for COVID-19 host genetic studies using nasopharyngeal samples from patients. The methods were compared regarding number of required steps for execution, sample handling time, quality and quantity of the extracted material and application in genetic studies. The Chelex(®)100 method was found to be cheapest (33 and 13 times cheaper than the commercial kit and phenol-chloroform, respectively), give the highest DNA yield (306 and 69 times higher than the commercial kit and phenol-chloroform, respectively), with the least handling steps while providing adequate DNA quality for downstream applications. Together, our results show that the Chelex(®)100 resin is an inexpensive, safe, simple, fast, and suitable method for DNA extraction of nasopharyngeal samples from COVID-19 patients for genetics studies. This is particularly relevant in developing countries where cost and handling are critical steps in material processing. Public Library of Science 2023-10-30 /pmc/articles/PMC10615309/ /pubmed/37903126 http://dx.doi.org/10.1371/journal.pone.0287551 Text en © 2023 Silva et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
da Silva, Ronaldo Celerino
de Lima, Suelen Cristina
dos Santos Reis, Wendell Palôma Maria
de Magalhães, Jurandy Júnior Ferraz
Magalhães, Ronaldo Nascimento de Oliveira
Rathi, Brijesh
Kohl, Alain
Bezerra, Marcos André Cavalcanti
Pena, Lindomar
Comparison of DNA extraction methods for COVID-19 host genetics studies
title Comparison of DNA extraction methods for COVID-19 host genetics studies
title_full Comparison of DNA extraction methods for COVID-19 host genetics studies
title_fullStr Comparison of DNA extraction methods for COVID-19 host genetics studies
title_full_unstemmed Comparison of DNA extraction methods for COVID-19 host genetics studies
title_short Comparison of DNA extraction methods for COVID-19 host genetics studies
title_sort comparison of dna extraction methods for covid-19 host genetics studies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10615309/
https://www.ncbi.nlm.nih.gov/pubmed/37903126
http://dx.doi.org/10.1371/journal.pone.0287551
work_keys_str_mv AT dasilvaronaldocelerino comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT delimasuelencristina comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT dossantosreiswendellpalomamaria comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT demagalhaesjurandyjuniorferraz comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT magalhaesronaldonascimentodeoliveira comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT rathibrijesh comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT kohlalain comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT bezerramarcosandrecavalcanti comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies
AT penalindomar comparisonofdnaextractionmethodsforcovid19hostgeneticsstudies