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One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses
Water contamination by viruses has an increasing worldwide impact on human health, and has led to requirements for accurate and quantitative molecular tools. Here, we report the first one-step reverse-transcription droplet digital PCR-based absolute quantification of a RNA virus (rotavirus) in diffe...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3892107/ https://www.ncbi.nlm.nih.gov/pubmed/24276251 http://dx.doi.org/10.1007/s00216-013-7476-y |
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author | Rački, Nejc Morisset, Dany Gutierrez-Aguirre, Ion Ravnikar, Maja |
author_facet | Rački, Nejc Morisset, Dany Gutierrez-Aguirre, Ion Ravnikar, Maja |
author_sort | Rački, Nejc |
collection | PubMed |
description | Water contamination by viruses has an increasing worldwide impact on human health, and has led to requirements for accurate and quantitative molecular tools. Here, we report the first one-step reverse-transcription droplet digital PCR-based absolute quantification of a RNA virus (rotavirus) in different types of surface water samples. This quantification method proved to be more precise and more tolerant to inhibitory substances than the benchmarking reverse-transcription real-time PCR (RT-qPCR), and needs no standard curve. This new tool is fully amenable for the quantification of viruses in the particularly low concentrations usually found in water samples. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-013-7476-y) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-3892107 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-38921072014-01-21 One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses Rački, Nejc Morisset, Dany Gutierrez-Aguirre, Ion Ravnikar, Maja Anal Bioanal Chem Rapid Communication Water contamination by viruses has an increasing worldwide impact on human health, and has led to requirements for accurate and quantitative molecular tools. Here, we report the first one-step reverse-transcription droplet digital PCR-based absolute quantification of a RNA virus (rotavirus) in different types of surface water samples. This quantification method proved to be more precise and more tolerant to inhibitory substances than the benchmarking reverse-transcription real-time PCR (RT-qPCR), and needs no standard curve. This new tool is fully amenable for the quantification of viruses in the particularly low concentrations usually found in water samples. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-013-7476-y) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2013-11-26 2014 /pmc/articles/PMC3892107/ /pubmed/24276251 http://dx.doi.org/10.1007/s00216-013-7476-y Text en © The Author(s) 2013 https://creativecommons.org/licenses/by/2.0/ Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Rapid Communication Rački, Nejc Morisset, Dany Gutierrez-Aguirre, Ion Ravnikar, Maja One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses |
title | One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses |
title_full | One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses |
title_fullStr | One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses |
title_full_unstemmed | One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses |
title_short | One-step RT-droplet digital PCR: a breakthrough in the quantification of waterborne RNA viruses |
title_sort | one-step rt-droplet digital pcr: a breakthrough in the quantification of waterborne rna viruses |
topic | Rapid Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3892107/ https://www.ncbi.nlm.nih.gov/pubmed/24276251 http://dx.doi.org/10.1007/s00216-013-7476-y |
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