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Increased sensitivity using real-time dPCR for detection of SARS-CoV-2

A real-time dPCR system was developed to improve the sensitivity, specificity and quantification accuracy of end point dPCR. We compared three technologies – real-time qPCR, end point dPCR and real-time dPCR – in the context of SARS-CoV-2. Some improvement in limit of detection was obtained with end...

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Autores principales: Duong, Kyra, Ou, Jiajia, Li, Zhaoliang, Lv, Zhaoqing, Dong, Hao, Hu, Tao, Zhang, Yunyun, Hanna, Ava, Gordon, Skyler, Crynen, Gogce, Head, Steven R, Ordoukhanian, Phillip, Wang, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7888512/
https://www.ncbi.nlm.nih.gov/pubmed/33222514
http://dx.doi.org/10.2144/btn-2020-0133
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author Duong, Kyra
Ou, Jiajia
Li, Zhaoliang
Lv, Zhaoqing
Dong, Hao
Hu, Tao
Zhang, Yunyun
Hanna, Ava
Gordon, Skyler
Crynen, Gogce
Head, Steven R
Ordoukhanian, Phillip
Wang, Yan
author_facet Duong, Kyra
Ou, Jiajia
Li, Zhaoliang
Lv, Zhaoqing
Dong, Hao
Hu, Tao
Zhang, Yunyun
Hanna, Ava
Gordon, Skyler
Crynen, Gogce
Head, Steven R
Ordoukhanian, Phillip
Wang, Yan
author_sort Duong, Kyra
collection PubMed
description A real-time dPCR system was developed to improve the sensitivity, specificity and quantification accuracy of end point dPCR. We compared three technologies – real-time qPCR, end point dPCR and real-time dPCR – in the context of SARS-CoV-2. Some improvement in limit of detection was obtained with end point dPCR compared with real-time qPCR, and the limit of detection was further improved with the newly developed real-time dPCR technology through removal of false-positive signals. Real-time dPCR showed increased linear dynamic range compared with end point dPCR based on quantitation from amplification curves. Real-time dPCR can improve the performance of TaqMan assays beyond real-time qPCR and end point dPCR with better sensitivity and specificity, absolute quantification and a wider linear range of detection.
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spelling pubmed-78885122021-02-26 Increased sensitivity using real-time dPCR for detection of SARS-CoV-2 Duong, Kyra Ou, Jiajia Li, Zhaoliang Lv, Zhaoqing Dong, Hao Hu, Tao Zhang, Yunyun Hanna, Ava Gordon, Skyler Crynen, Gogce Head, Steven R Ordoukhanian, Phillip Wang, Yan Biotechniques Reports A real-time dPCR system was developed to improve the sensitivity, specificity and quantification accuracy of end point dPCR. We compared three technologies – real-time qPCR, end point dPCR and real-time dPCR – in the context of SARS-CoV-2. Some improvement in limit of detection was obtained with end point dPCR compared with real-time qPCR, and the limit of detection was further improved with the newly developed real-time dPCR technology through removal of false-positive signals. Real-time dPCR showed increased linear dynamic range compared with end point dPCR based on quantitation from amplification curves. Real-time dPCR can improve the performance of TaqMan assays beyond real-time qPCR and end point dPCR with better sensitivity and specificity, absolute quantification and a wider linear range of detection. Future Science Ltd 2020-11-23 2020-10 /pmc/articles/PMC7888512/ /pubmed/33222514 http://dx.doi.org/10.2144/btn-2020-0133 Text en © 2020 Yan Wang This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Reports
Duong, Kyra
Ou, Jiajia
Li, Zhaoliang
Lv, Zhaoqing
Dong, Hao
Hu, Tao
Zhang, Yunyun
Hanna, Ava
Gordon, Skyler
Crynen, Gogce
Head, Steven R
Ordoukhanian, Phillip
Wang, Yan
Increased sensitivity using real-time dPCR for detection of SARS-CoV-2
title Increased sensitivity using real-time dPCR for detection of SARS-CoV-2
title_full Increased sensitivity using real-time dPCR for detection of SARS-CoV-2
title_fullStr Increased sensitivity using real-time dPCR for detection of SARS-CoV-2
title_full_unstemmed Increased sensitivity using real-time dPCR for detection of SARS-CoV-2
title_short Increased sensitivity using real-time dPCR for detection of SARS-CoV-2
title_sort increased sensitivity using real-time dpcr for detection of sars-cov-2
topic Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7888512/
https://www.ncbi.nlm.nih.gov/pubmed/33222514
http://dx.doi.org/10.2144/btn-2020-0133
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