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Digital RT-PCR Chip method for detection of SARS-CoV-2 virus

The “gold standard” method for detection of SARS-CoV-2 is the real time reverse transcription-polymerase chain reaction, but due to pre-analytical and technical limitations, biological samples with low viral load are not sometimes detected. For this purpose a digital RT-PCR method on-chip was develo...

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Autores principales: Dioni, Laura, Orlandi, Annarosa, Uceda Renteria, Sara, Favero, Chiara, Solazzo, Giulia, Oggioni, Massimo, Bollati, Valentina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9383957/
https://www.ncbi.nlm.nih.gov/pubmed/35985558
http://dx.doi.org/10.1016/j.jim.2022.113339
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author Dioni, Laura
Orlandi, Annarosa
Uceda Renteria, Sara
Favero, Chiara
Solazzo, Giulia
Oggioni, Massimo
Bollati, Valentina
author_facet Dioni, Laura
Orlandi, Annarosa
Uceda Renteria, Sara
Favero, Chiara
Solazzo, Giulia
Oggioni, Massimo
Bollati, Valentina
author_sort Dioni, Laura
collection PubMed
description The “gold standard” method for detection of SARS-CoV-2 is the real time reverse transcription-polymerase chain reaction, but due to pre-analytical and technical limitations, biological samples with low viral load are not sometimes detected. For this purpose a digital RT-PCR method on-chip was developed for detection of the SARS-CoV-2 virus, using two TaqMan™ Assays for quantification of the N Protein (Nucleocapsid) and the S Protein (Spike), and the QuantStudio™ 3D Digital PCR instrument. The method was applied to assess the nasopharyngeal swabs of asymptomatic subjects recruited in the UNICORN Study. The digital RT-PCR method is characterized by a higher sensitivity than the RT-qPCR method, even if performed with the same TaqMan™, and could be a promising tool for SARS-CoV-2 viral load quantification.
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spelling pubmed-93839572022-08-17 Digital RT-PCR Chip method for detection of SARS-CoV-2 virus Dioni, Laura Orlandi, Annarosa Uceda Renteria, Sara Favero, Chiara Solazzo, Giulia Oggioni, Massimo Bollati, Valentina J Immunol Methods Technical Note The “gold standard” method for detection of SARS-CoV-2 is the real time reverse transcription-polymerase chain reaction, but due to pre-analytical and technical limitations, biological samples with low viral load are not sometimes detected. For this purpose a digital RT-PCR method on-chip was developed for detection of the SARS-CoV-2 virus, using two TaqMan™ Assays for quantification of the N Protein (Nucleocapsid) and the S Protein (Spike), and the QuantStudio™ 3D Digital PCR instrument. The method was applied to assess the nasopharyngeal swabs of asymptomatic subjects recruited in the UNICORN Study. The digital RT-PCR method is characterized by a higher sensitivity than the RT-qPCR method, even if performed with the same TaqMan™, and could be a promising tool for SARS-CoV-2 viral load quantification. Elsevier B.V. 2022-10 2022-08-17 /pmc/articles/PMC9383957/ /pubmed/35985558 http://dx.doi.org/10.1016/j.jim.2022.113339 Text en © 2022 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 Technical Note
Dioni, Laura
Orlandi, Annarosa
Uceda Renteria, Sara
Favero, Chiara
Solazzo, Giulia
Oggioni, Massimo
Bollati, Valentina
Digital RT-PCR Chip method for detection of SARS-CoV-2 virus
title Digital RT-PCR Chip method for detection of SARS-CoV-2 virus
title_full Digital RT-PCR Chip method for detection of SARS-CoV-2 virus
title_fullStr Digital RT-PCR Chip method for detection of SARS-CoV-2 virus
title_full_unstemmed Digital RT-PCR Chip method for detection of SARS-CoV-2 virus
title_short Digital RT-PCR Chip method for detection of SARS-CoV-2 virus
title_sort digital rt-pcr chip method for detection of sars-cov-2 virus
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9383957/
https://www.ncbi.nlm.nih.gov/pubmed/35985558
http://dx.doi.org/10.1016/j.jim.2022.113339
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