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Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR

BACKGROUND: Research efforts for the management of cancer, in particular for lung cancer, are directed to identify new strategies for its early detection. MicroRNAs (miRNAs) are a new promising class of circulating biomarkers for cancer detection, but lack of consensus on data normalization methods...

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Autores principales: Conte, Davide, Verri, Carla, Borzi, Cristina, Suatoni, Paola, Pastorino, Ugo, Sozzi, Gabriella, Fortunato, Orazio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619272/
https://www.ncbi.nlm.nih.gov/pubmed/26493562
http://dx.doi.org/10.1186/s12864-015-2097-9
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author Conte, Davide
Verri, Carla
Borzi, Cristina
Suatoni, Paola
Pastorino, Ugo
Sozzi, Gabriella
Fortunato, Orazio
author_facet Conte, Davide
Verri, Carla
Borzi, Cristina
Suatoni, Paola
Pastorino, Ugo
Sozzi, Gabriella
Fortunato, Orazio
author_sort Conte, Davide
collection PubMed
description BACKGROUND: Research efforts for the management of cancer, in particular for lung cancer, are directed to identify new strategies for its early detection. MicroRNAs (miRNAs) are a new promising class of circulating biomarkers for cancer detection, but lack of consensus on data normalization methods has affected the diagnostic potential of circulating miRNAs. There is a growing interest in techniques that allow an absolute quantification of miRNAs which could be useful for early diagnosis. Recently, digital PCR, mainly based on droplets generation, emerged as an affordable technology for precise and absolute quantification of nucleic acids. RESULTS: In this work, we described a new interesting approach for profiling circulating miRNAs in plasma samples using a chip-based platform, the QuantStudio 3D digital PCR. The proposed method was validated using synthethic oligonucleotide at serial dilutions in plasma samples of lung cancer patients and in lung tissues and cell lines. CONCLUSION: Given its reproducibility and reliability, our approach could be potentially applied for the identification and quantification of miRNAs in other biological samples such as circulating exosomes or protein complexes. As chip-digital PCR becomes more established, it would be a robust tool for quantitative assessment of miRNA copy number for diagnosis of lung cancer and other diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-2097-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-46192722015-10-26 Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR Conte, Davide Verri, Carla Borzi, Cristina Suatoni, Paola Pastorino, Ugo Sozzi, Gabriella Fortunato, Orazio BMC Genomics Methodology Article BACKGROUND: Research efforts for the management of cancer, in particular for lung cancer, are directed to identify new strategies for its early detection. MicroRNAs (miRNAs) are a new promising class of circulating biomarkers for cancer detection, but lack of consensus on data normalization methods has affected the diagnostic potential of circulating miRNAs. There is a growing interest in techniques that allow an absolute quantification of miRNAs which could be useful for early diagnosis. Recently, digital PCR, mainly based on droplets generation, emerged as an affordable technology for precise and absolute quantification of nucleic acids. RESULTS: In this work, we described a new interesting approach for profiling circulating miRNAs in plasma samples using a chip-based platform, the QuantStudio 3D digital PCR. The proposed method was validated using synthethic oligonucleotide at serial dilutions in plasma samples of lung cancer patients and in lung tissues and cell lines. CONCLUSION: Given its reproducibility and reliability, our approach could be potentially applied for the identification and quantification of miRNAs in other biological samples such as circulating exosomes or protein complexes. As chip-digital PCR becomes more established, it would be a robust tool for quantitative assessment of miRNA copy number for diagnosis of lung cancer and other diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-2097-9) contains supplementary material, which is available to authorized users. BioMed Central 2015-10-23 /pmc/articles/PMC4619272/ /pubmed/26493562 http://dx.doi.org/10.1186/s12864-015-2097-9 Text en © Conte et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Conte, Davide
Verri, Carla
Borzi, Cristina
Suatoni, Paola
Pastorino, Ugo
Sozzi, Gabriella
Fortunato, Orazio
Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR
title Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR
title_full Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR
title_fullStr Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR
title_full_unstemmed Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR
title_short Novel method to detect microRNAs using chip-based QuantStudio 3D digital PCR
title_sort novel method to detect micrornas using chip-based quantstudio 3d digital pcr
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619272/
https://www.ncbi.nlm.nih.gov/pubmed/26493562
http://dx.doi.org/10.1186/s12864-015-2097-9
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