Cargando…

A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans

Circulating microRNAs (miRNAs) have become increasingly popular biomarker candidates in various diseases. However, heparin-based anticoagulants might affect the detection of target miRNAs in blood samples during quantitative polymerase chain reaction (qPCR)-based analysis of miRNAs involving RNA ext...

Descripción completa

Detalles Bibliográficos
Autores principales: Nilsson, Abraham, Nerhall, Anna Maria, Vechetti, Ivan, Fornander, Lotta, Wiklund, Simon, Alkner, Björn, Schilcher, Jörg, von Walden, Ferdinand
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9632808/
https://www.ncbi.nlm.nih.gov/pubmed/36327247
http://dx.doi.org/10.1371/journal.pone.0277008
_version_ 1784824116918878208
author Nilsson, Abraham
Nerhall, Anna Maria
Vechetti, Ivan
Fornander, Lotta
Wiklund, Simon
Alkner, Björn
Schilcher, Jörg
von Walden, Ferdinand
author_facet Nilsson, Abraham
Nerhall, Anna Maria
Vechetti, Ivan
Fornander, Lotta
Wiklund, Simon
Alkner, Björn
Schilcher, Jörg
von Walden, Ferdinand
author_sort Nilsson, Abraham
collection PubMed
description Circulating microRNAs (miRNAs) have become increasingly popular biomarker candidates in various diseases. However, heparin-based anticoagulants might affect the detection of target miRNAs in blood samples during quantitative polymerase chain reaction (qPCR)-based analysis of miRNAs involving RNA extraction, cDNA synthesis and the polymerase catalyzed reaction. Because low-molecular-weight heparins (LMWH) are widely used in routine healthcare, we aimed to investigate whether a prophylactic dose of the LMWH tinzaparin influences qPCR-based quantification of circulating miRNAs. A total of 30 subjects were included: 16 fracture patients with tinzaparin treatment and 14 non-fracture controls without anticoagulation therapy. To control for the effect of tinzaparin on miRNA analysis an identical concentration of synthetic miRNAs was added to plasma, isolated RNA and prepared complementary DNA (cDNA) from all samples in both groups. No significant difference was observed for cDNA synthesis or qPCR when comparing tinzaparin-treated patients with untreated controls. Among the tinzaparin-treated patients, plasma levels of six endogenous miRNAs (hsa-let-7i-5p, hsa-miR-30e-5p, hsa-miR-222-3p, hsa-miR-1-3p, hsa-miR-133a-3p, hsa-miR-133b) were measured before and one to six hours after a subcutaneous injection of tinzaparin 4500IU. No significant effect was observed for any of the investigated miRNAs. A prophylactic dose of 4500IU tinzaparin does not seem to affect cDNA synthesis or qRT-PCR-based quantification of circulating miRNAs.
format Online
Article
Text
id pubmed-9632808
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-96328082022-11-04 A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans Nilsson, Abraham Nerhall, Anna Maria Vechetti, Ivan Fornander, Lotta Wiklund, Simon Alkner, Björn Schilcher, Jörg von Walden, Ferdinand PLoS One Research Article Circulating microRNAs (miRNAs) have become increasingly popular biomarker candidates in various diseases. However, heparin-based anticoagulants might affect the detection of target miRNAs in blood samples during quantitative polymerase chain reaction (qPCR)-based analysis of miRNAs involving RNA extraction, cDNA synthesis and the polymerase catalyzed reaction. Because low-molecular-weight heparins (LMWH) are widely used in routine healthcare, we aimed to investigate whether a prophylactic dose of the LMWH tinzaparin influences qPCR-based quantification of circulating miRNAs. A total of 30 subjects were included: 16 fracture patients with tinzaparin treatment and 14 non-fracture controls without anticoagulation therapy. To control for the effect of tinzaparin on miRNA analysis an identical concentration of synthetic miRNAs was added to plasma, isolated RNA and prepared complementary DNA (cDNA) from all samples in both groups. No significant difference was observed for cDNA synthesis or qPCR when comparing tinzaparin-treated patients with untreated controls. Among the tinzaparin-treated patients, plasma levels of six endogenous miRNAs (hsa-let-7i-5p, hsa-miR-30e-5p, hsa-miR-222-3p, hsa-miR-1-3p, hsa-miR-133a-3p, hsa-miR-133b) were measured before and one to six hours after a subcutaneous injection of tinzaparin 4500IU. No significant effect was observed for any of the investigated miRNAs. A prophylactic dose of 4500IU tinzaparin does not seem to affect cDNA synthesis or qRT-PCR-based quantification of circulating miRNAs. Public Library of Science 2022-11-03 /pmc/articles/PMC9632808/ /pubmed/36327247 http://dx.doi.org/10.1371/journal.pone.0277008 Text en © 2022 Nilsson et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Nilsson, Abraham
Nerhall, Anna Maria
Vechetti, Ivan
Fornander, Lotta
Wiklund, Simon
Alkner, Björn
Schilcher, Jörg
von Walden, Ferdinand
A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans
title A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans
title_full A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans
title_fullStr A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans
title_full_unstemmed A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans
title_short A prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qPCR-based assessment of circulating levels of miRNA in humans
title_sort prophylactic subcutaneous dose of the anticoagulant tinzaparin does not influence qpcr-based assessment of circulating levels of mirna in humans
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9632808/
https://www.ncbi.nlm.nih.gov/pubmed/36327247
http://dx.doi.org/10.1371/journal.pone.0277008
work_keys_str_mv AT nilssonabraham aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT nerhallannamaria aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT vechettiivan aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT fornanderlotta aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT wiklundsimon aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT alknerbjorn aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT schilcherjorg aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT vonwaldenferdinand aprophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT nilssonabraham prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT nerhallannamaria prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT vechettiivan prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT fornanderlotta prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT wiklundsimon prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT alknerbjorn prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT schilcherjorg prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans
AT vonwaldenferdinand prophylacticsubcutaneousdoseoftheanticoagulanttinzaparindoesnotinfluenceqpcrbasedassessmentofcirculatinglevelsofmirnainhumans