Cargando…
Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers
In December 2019, the Chinese Center for Disease Control (CDC of China) reported an outbreak of pneumonia in the city of Wuhan (Hubei province, China) that haunted the world, resulting in a global pandemic. This outbreak was caused by a betacoronavirus named severe acute respiratory syndrome coronav...
Autores principales: | , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Author(s). Published by Elsevier Ltd.
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9891106/ https://www.ncbi.nlm.nih.gov/pubmed/36742065 http://dx.doi.org/10.1016/j.jcvp.2023.100134 |
_version_ | 1784881077267988480 |
---|---|
author | Barboza, Victor dos Santos Domingues, William Borges de Souza, Thobias Toniolo Collares, Tiago Veiras Seixas, Fabiana Kommling Pacheco, Bruna Silveira Sousa, Fernanda Severo Sabedra Oliveira, Thaís Larré de Lima, Marcelo de Pereira, Claúdio Martin Pereira Spilki, Fernando Rosado Giongo, Janice Luehring Vaucher, Rodrigo de Almeida |
author_facet | Barboza, Victor dos Santos Domingues, William Borges de Souza, Thobias Toniolo Collares, Tiago Veiras Seixas, Fabiana Kommling Pacheco, Bruna Silveira Sousa, Fernanda Severo Sabedra Oliveira, Thaís Larré de Lima, Marcelo de Pereira, Claúdio Martin Pereira Spilki, Fernando Rosado Giongo, Janice Luehring Vaucher, Rodrigo de Almeida |
author_sort | Barboza, Victor dos Santos |
collection | PubMed |
description | In December 2019, the Chinese Center for Disease Control (CDC of China) reported an outbreak of pneumonia in the city of Wuhan (Hubei province, China) that haunted the world, resulting in a global pandemic. This outbreak was caused by a betacoronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Several of these cases have been observed in healthcare professionals working in hospitals and providing care on the pandemic's frontline. In the present study, nasopharyngeal swab samples of healthcare workers were used to assess the performance of the reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay and subsequently compared with the real‐time reverse-transcription quantitative PCR (RT-qPCR) method. Thus, in this study, we validated a method for detecting SARS-CoV-2 based on RT-LAMP that can be used to diagnose these workers. The methodology used was based on analyzing the sensitivity, specificity, evaluation of the detection limit, and cross-reaction with other respiratory viruses. The agreement was estimated using a dispersion diagram designed using the Bland-Altman method. A total of 100 clinical specimens of nasopharyngeal swabs were collected from symptomatic and asymptomatic healthcare workers in Pelotas, Brazil, during the SARS-CoV-2 outbreak. RT-LAMP assay, it was possible to detect SARS-CoV-2 in 96.7% of the healthcare professionals tested using the E gene and N gene primers approximately and 100% for the gene of human β-actin. The observed agreement was considered excellent for the primer set of the E and N genes (k = 0.957 and k = 0.896), respectively. The sensitivity of the RT-LAMP assay was positive for the primer set of the E gene, detected to approximately 2 copies per reaction. For the primer set of the N gene, the assay was possible to verify an LoD of approximately 253 copies per reaction. After executing the RT-LAMP assay, no positive reactions were observed for any of the virus respiratory tested. Therefore, we conclude that RT-LAMP is effective for rapid molecular diagnosis during the COVID-19 outbreak period in healthcare professionals. |
format | Online Article Text |
id | pubmed-9891106 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The Author(s). Published by Elsevier Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-98911062023-02-01 Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers Barboza, Victor dos Santos Domingues, William Borges de Souza, Thobias Toniolo Collares, Tiago Veiras Seixas, Fabiana Kommling Pacheco, Bruna Silveira Sousa, Fernanda Severo Sabedra Oliveira, Thaís Larré de Lima, Marcelo de Pereira, Claúdio Martin Pereira Spilki, Fernando Rosado Giongo, Janice Luehring Vaucher, Rodrigo de Almeida J Clin Virol Plus Article In December 2019, the Chinese Center for Disease Control (CDC of China) reported an outbreak of pneumonia in the city of Wuhan (Hubei province, China) that haunted the world, resulting in a global pandemic. This outbreak was caused by a betacoronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Several of these cases have been observed in healthcare professionals working in hospitals and providing care on the pandemic's frontline. In the present study, nasopharyngeal swab samples of healthcare workers were used to assess the performance of the reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay and subsequently compared with the real‐time reverse-transcription quantitative PCR (RT-qPCR) method. Thus, in this study, we validated a method for detecting SARS-CoV-2 based on RT-LAMP that can be used to diagnose these workers. The methodology used was based on analyzing the sensitivity, specificity, evaluation of the detection limit, and cross-reaction with other respiratory viruses. The agreement was estimated using a dispersion diagram designed using the Bland-Altman method. A total of 100 clinical specimens of nasopharyngeal swabs were collected from symptomatic and asymptomatic healthcare workers in Pelotas, Brazil, during the SARS-CoV-2 outbreak. RT-LAMP assay, it was possible to detect SARS-CoV-2 in 96.7% of the healthcare professionals tested using the E gene and N gene primers approximately and 100% for the gene of human β-actin. The observed agreement was considered excellent for the primer set of the E and N genes (k = 0.957 and k = 0.896), respectively. The sensitivity of the RT-LAMP assay was positive for the primer set of the E gene, detected to approximately 2 copies per reaction. For the primer set of the N gene, the assay was possible to verify an LoD of approximately 253 copies per reaction. After executing the RT-LAMP assay, no positive reactions were observed for any of the virus respiratory tested. Therefore, we conclude that RT-LAMP is effective for rapid molecular diagnosis during the COVID-19 outbreak period in healthcare professionals. The Author(s). Published by Elsevier Ltd. 2023-06 2023-01-27 /pmc/articles/PMC9891106/ /pubmed/36742065 http://dx.doi.org/10.1016/j.jcvp.2023.100134 Text en © 2023 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 Barboza, Victor dos Santos Domingues, William Borges de Souza, Thobias Toniolo Collares, Tiago Veiras Seixas, Fabiana Kommling Pacheco, Bruna Silveira Sousa, Fernanda Severo Sabedra Oliveira, Thaís Larré de Lima, Marcelo de Pereira, Claúdio Martin Pereira Spilki, Fernando Rosado Giongo, Janice Luehring Vaucher, Rodrigo de Almeida Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers |
title | Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers |
title_full | Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers |
title_fullStr | Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers |
title_full_unstemmed | Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers |
title_short | Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers |
title_sort | reverse transcription-loop-mediated isothermal amplification (rt-lamp) assay as a rapid molecular diagnostic tool for covid-19 in healthcare workers |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9891106/ https://www.ncbi.nlm.nih.gov/pubmed/36742065 http://dx.doi.org/10.1016/j.jcvp.2023.100134 |
work_keys_str_mv | AT barbozavictordossantos reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT domingueswilliamborges reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT desouzathobiastoniolo reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT collarestiagoveiras reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT seixasfabianakommling reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT pachecobrunasilveira reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT sousafernandaseverosabedra reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT oliveirathaislarre reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT delimamarcelo reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT depereiraclaudiomartinpereira reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT spilkifernandorosado reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT giongojaniceluehring reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers AT vaucherrodrigodealmeida reversetranscriptionloopmediatedisothermalamplificationrtlampassayasarapidmoleculardiagnostictoolforcovid19inhealthcareworkers |