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Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies

In the last decade, circulating nucleic acids such as microRNAs (miRNAs) and cell-free DNA (cfDNA) have become increasingly important in serving as potential novel biomarkers for a variety of human diseases. If cell-free nucleic acids are to become routinely used in diagnostics, the difference in pl...

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Autores principales: Ward Gahlawat, Aoife, Lenhardt, Judith, Witte, Tania, Keitel, Denise, Kaufhold, Anna, Maass, Kendra K, Pajtler, Kristian W, Sohn, Christof, Schott, Sarah
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387045/
https://www.ncbi.nlm.nih.gov/pubmed/30736351
http://dx.doi.org/10.3390/ijms20030704
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author Ward Gahlawat, Aoife
Lenhardt, Judith
Witte, Tania
Keitel, Denise
Kaufhold, Anna
Maass, Kendra K
Pajtler, Kristian W
Sohn, Christof
Schott, Sarah
author_facet Ward Gahlawat, Aoife
Lenhardt, Judith
Witte, Tania
Keitel, Denise
Kaufhold, Anna
Maass, Kendra K
Pajtler, Kristian W
Sohn, Christof
Schott, Sarah
author_sort Ward Gahlawat, Aoife
collection PubMed
description In the last decade, circulating nucleic acids such as microRNAs (miRNAs) and cell-free DNA (cfDNA) have become increasingly important in serving as potential novel biomarkers for a variety of human diseases. If cell-free nucleic acids are to become routinely used in diagnostics, the difference in plasma miRNA and cfDNA levels between healthy and diseased subjects must exceed pre-analytical and analytical variability. Until now, few studies have addressed the time limitations of pre-processing or explored the potential use of long-term blood storage tubes, which might need to be implemented in real-life diagnostics. In this study, we analyzed the stability of four breast cancer-associated miRNAs and two cancer-associated genes under various storage conditions, to test their limitations for potential application in clinical diagnostics. In two consecutive experiments, we tested the limits of conventional EDTA tubes, as well as long-term storage blood collection tubes (BCTs) from four different manufacturers. We found that circulating miRNAs are relatively stable when stored in EDTA monovettes for up to 12 h before processing. When stored in BCTs, circulating miRNAs and cfDNA are stable for up to 7 days, depending on the manufacturer. Norgen tubes were superior for cfDNA yield, while Streck tubes performed the worst in our study with hemolysis induction. In conclusion, plasma prepared from whole blood is suitable for the quantification of both cf-miRNAs and cfDNA simultaneously.
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spelling pubmed-63870452019-02-27 Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies Ward Gahlawat, Aoife Lenhardt, Judith Witte, Tania Keitel, Denise Kaufhold, Anna Maass, Kendra K Pajtler, Kristian W Sohn, Christof Schott, Sarah Int J Mol Sci Communication In the last decade, circulating nucleic acids such as microRNAs (miRNAs) and cell-free DNA (cfDNA) have become increasingly important in serving as potential novel biomarkers for a variety of human diseases. If cell-free nucleic acids are to become routinely used in diagnostics, the difference in plasma miRNA and cfDNA levels between healthy and diseased subjects must exceed pre-analytical and analytical variability. Until now, few studies have addressed the time limitations of pre-processing or explored the potential use of long-term blood storage tubes, which might need to be implemented in real-life diagnostics. In this study, we analyzed the stability of four breast cancer-associated miRNAs and two cancer-associated genes under various storage conditions, to test their limitations for potential application in clinical diagnostics. In two consecutive experiments, we tested the limits of conventional EDTA tubes, as well as long-term storage blood collection tubes (BCTs) from four different manufacturers. We found that circulating miRNAs are relatively stable when stored in EDTA monovettes for up to 12 h before processing. When stored in BCTs, circulating miRNAs and cfDNA are stable for up to 7 days, depending on the manufacturer. Norgen tubes were superior for cfDNA yield, while Streck tubes performed the worst in our study with hemolysis induction. In conclusion, plasma prepared from whole blood is suitable for the quantification of both cf-miRNAs and cfDNA simultaneously. MDPI 2019-02-06 /pmc/articles/PMC6387045/ /pubmed/30736351 http://dx.doi.org/10.3390/ijms20030704 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Ward Gahlawat, Aoife
Lenhardt, Judith
Witte, Tania
Keitel, Denise
Kaufhold, Anna
Maass, Kendra K
Pajtler, Kristian W
Sohn, Christof
Schott, Sarah
Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies
title Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies
title_full Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies
title_fullStr Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies
title_full_unstemmed Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies
title_short Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies
title_sort evaluation of storage tubes for combined analysis of circulating nucleic acids in liquid biopsies
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387045/
https://www.ncbi.nlm.nih.gov/pubmed/30736351
http://dx.doi.org/10.3390/ijms20030704
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