ArticleBiomolecular detection and quantification2016
Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs.
Article in Biomolecular detection and quantification, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 35 citations in OpenAlex.
- Exploring the link between fatty acids, FADS2 expression, and ADHD using structural equation modeling.European child & adolescent psychiatry · 2026Article
- Heparinase I treatment to overcome RNA quantification interference in heparinized liver donor samples: One size fits all?PloS one · 2025Observational
- Recommendations for detection, validation, and evaluation of RNA editing events in cardiovascular and neurological/neurodegenerative diseases.Molecular therapy. Nucleic acids · 2024Review
- Biological basis of extensive pleiotropy between blood traits and cancer risk.Genome medicine · 2024Article
- A Simple, Rapid, and Effective Heparinase Protocol to Enable Nucleic Acid Study from Frozen Heparinized Plasma.Methods and protocols · 2023Article
- Plasma microrna quantification protocol.Vessel plus · 2023Article
- Circulating microRNAs as Reliable Tumor Biomarkers: Opportunities and Challenges Facing Clinical Application.The Journal of pharmacology and experimental therapeutics · 2023Review
- Hydrogen sulfide and miR21 are suitable biomarkers of hypoxic exposure.American journal of physiology. Regulatory, integrative and comparative physiology · 2022Article
- Relationship between the Levels of lncRNA H19 in Plasma and Different Adipose Tissue Depots with Patients' Response to Bariatric Surgery.Life (Basel, Switzerland) · 2022Article
- Tools for Assessing Translation inJournal of fungi (Basel, Switzerland) · 2021Review
- Different Expressions of Pericardial Fluid MicroRNAs in Patients With Arrhythmogenic Right Ventricular Cardiomyopathy and Ischemic Heart Disease Undergoing Ventricular Tachycardia Ablation.Frontiers in cardiovascular medicine · 2021Article
- A simplified protocol for profiling heparin-contaminated circulating miRNAs: by microfluidic array.Molecular biology reports · 2020Article
- Practical Procedures for Improving Detection of Circulating miRNAs in Cardiovascular Diseases.Journal of cardiovascular translational research · 2020Article
- Circulating microRNAs as possible biomarkers for coronary artery disease: a narrative review.EJIFCC · 2019Review
- Release of Mitochondrial and Nuclear DNA During On-Pump Heart Surgery: Kinetics and Relation to Extracellular Vesicles.Journal of cardiovascular translational research · 2019Article
- Mapping of genomic EGFRvIII deletions in glioblastoma: insight into rearrangement mechanisms and biomarker development.Neuro-oncology · 2018Article
- Establishment of an easy and straight forward heparinase protocol to analyse circulating and myocardial tissue micro-RNA during coronary artery-bypass-graft surgery.Scientific reports · 2018Article
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8 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundmicroRNAs have recently been identified as powerful biomarkers of human disease. Reliable polymerase chain reaction (PCR)-based quantification of nucleic acids in clinical samples contaminated with polymerase inhibitor heparin requires deheparinization. However, the effects of deheparinization procedure on quantification of nucleic acids remain largely unknown. The aim of this study was to determine whether the deheparinization procedure completely eliminates the inhibition of amplification, while maintaining RNA integrity and technical variability of the measured microRNA levels.
methodsHeparinized plasma from 9 patients undergoing coronary artery bypass grafting (CABG) and the heparin-free plasma from 58 rats were spiked with a synthetic RNA oligonucleotide and total RNA was extracted. The RNA solutions were then treated with heparinase I to remove contaminating heparin prior to reverse transcription. Levels of synthetic spike-in RNA oligonucleotide, as well as endogenous hsa-miR-1-3p and hsa-miR-208a-3p, were measured using quantitative reverse transcription PCR (RT-qPCR). The amplification efficiency and presence of inhibitors in individual samples were directly determined using calibration curves.
resultsIn contrast to RNA samples from rat plasma, RNA samples derived from the CABG patient plasma contained inhibitors, which were completely eliminated by treatment with heparinase. The procedure caused a decrease in the amount of detected RNA; however, the technical variability of the measured targets did not change, allowing for the quantification of circulating endogenous hsa-miR-1-3p and hsa-miR-208a-3p in the plasma of CABG patients.
conclusionsThe heparinase treatment procedure enables utilization of RT-qPCR for reliable microRNA quantification in heparinized plasma.
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