ArticleBiomarker research2014
Circumventing qPCR inhibition to amplify miRNAs in plasma.
Article in Biomarker research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers, 1 of them a synthesis that pooled it.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
19 citing papers in PubMed, 1 synthesis or guideline pooled it, 35 citations in OpenAlex.
- Biomarkers in Non-Small Cell Lung Cancers: Indian Consensus Guidelines for Molecular Testing.Advances in therapy · 2019Guideline
- Circulating microRNA profiles predict the severity of COVID-19 in hospitalized patients.Translational research : the journal of laboratory and clinical medicine · 2021Trial
- Circulating levels of miR-20b-5p are associated with survival in cardiogenic shock.Journal of molecular and cellular cardiology plus · 2025Article
- Innovative Applications and Challenges of Isothermal Amplification Technology in miRNA Detection.Current genomics · 2025Review
- Heparinase I treatment to overcome RNA quantification interference in heparinized liver donor samples: One size fits all?PloS one · 2025Observational
- Circulating microRNAs targeting coagulation and fibrinolysis in patients with severe COVID-19.Thrombosis journal · 2024Article
- A Simple, Rapid, and Effective Heparinase Protocol to Enable Nucleic Acid Study from Frozen Heparinized Plasma.Methods and protocols · 2023Article
- Evaluation of RNA isolation methods for microRNA quantification in a range of clinical biofluids.BMC biotechnology · 2021Article
- Peripheral blood microRNAs and the COVID-19 patient: methodological considerations, technical challenges and practice points.RNA biology · 2021Review
- MiRNA-15b and miRNA-125b are associated with regional Aβ-PET and FDG-PET uptake in cognitively normal individuals with subjective memory complaints.Translational psychiatry · 2021Article
- Extracellular Vesicles as Biomarkers of Acute Graft-vs.-Host Disease After Haploidentical Stem Cell Transplantation and Post-Transplant Cyclophosphamide.Frontiers in immunology · 2021Article
- A simplified protocol for profiling heparin-contaminated circulating miRNAs: by microfluidic array.Molecular biology reports · 2020Article
- Article
- Enzyme-linked oligonucleotide hybridization assay for direct oligo measurement in blood.Biology methods & protocols · 2019Article
- Sources and Functions of Extracellular Small RNAs in Human Circulation.Annual review of nutrition · 2016Review
- Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs.Biomolecular detection and quantification · 2016Article
- Optimized Collection Protocol for Plasma MicroRNA Measurement in Patients with Cardiovascular Disease.BioMed research international · 2016Article
- A microRNA profile associated with Opisthorchis viverrini-induced cholangiocarcinoma in tissue and plasma.BMC cancer · 2015Article
- Maternal pre-pregnancy body mass index and circulating microRNAs in pregnancy.Obesity research & clinical practiceArticle
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundCirculating microRNAs (c-miRNAs) have be identified in saliva, urine and blood, which has led to increasing interest in their development as biomarkers for diverse diseases including cancers. One of the key advantages of c-miRNAs over other biomarkers is the ability to be amplified and quantified by quantitative PCR (qPCR). However, at phlebotomy when whole blood is dispensed into heparinized tubes, residual levels of the anti-coagulant lithium heparin may remain in the plasma and hence with RNA isolated from the plasma. This can confound the detection of c-miRNAs by qPCR because it inhibits reverse transcriptase (RT). Here we present a procedure, modified from earlier techniques, to detect c-miRNAs in plasma that improves sensitivity and streamlines performance.
findingsTreatment of total RNA isolated from human blood plasma with Bacteroides heparinase I during reverse transcription at 37°C for one hour improved sensitivity and performance of the qPCR. This is in comparison to no treatment or treatment of the RNA prior to RT, which is the current suggested method and exposes plasma to Flavobacterium heparinum heparinase I for up to 2 hours before RT. This modest alteration improved qPCR performance and resulted in lowered threshold cycles (Ct) for detection of the target sequence, candidate c-miRNA biomarkers, and controls. It also reduced the expense and number of processing steps, shortening the duration of the assay and minimizing exposure of RNA to elevated temperatures.
conclusionIncorporating Bacteroides heparinase I treatment into conventional RT protocols targeting c-miRNA in plasma can be expected to expedite the discovery of biomarkers.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.