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Updated: May 3, 2026
Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR
Published on: June 26, 2016
Methodological challenges in utilizing miRNAs as circulating biomarkers
Leni Moldovan1, Kara E Batte, Joanne Trgovcich
1Division of Pulmonary, Allergy, Critical Care, Sleep Medicine, College of Medicine, The Ohio State University, Columbus, OH, USA.
Abstract:
MicroRNAs (miRNAs) have emerged as important regulators in the post-transcriptional control of gene expression. The discovery of their presence not only in tissues but also in extratissular fluids, including blood, urine and cerebro-spinal fluid, together with their changes in expression in various pathological conditions, has implicated these extracellular miRNAs as informative biomarkers of disease. However, exploiting miRNAs in this capacity requires methodological rigour. Here, we report several key procedural aspects of miRNA isolation from plasma and serum, as exemplified by research in cardiovascular and pulmonary diseases. We also highlight the advantages and disadvantages of various profiling methods to determine the expression levels of plasma- and serum-derived miRNAs. Attention to such methodological details is critical, as circulating miRNAs become diagnostic tools for various human diseases.
Insights
Extracellular microRNAs (miRNAs) in blood are promising disease biomarkers. Rigorous methods for isolating and profiling these circulating miRNAs are crucial for accurate diagnostics, particularly in cardiovascular and pulmonary diseases.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are key post-transcriptional gene regulators.
- Extracellular miRNAs found in bodily fluids like blood are implicated as disease biomarkers.
- Standardized methodologies are needed for reliable biomarker discovery.
Purpose of the Study:
- To detail critical procedural aspects of miRNA isolation from plasma and serum.
- To evaluate various profiling methods for quantifying circulating miRNAs.
- To emphasize methodological rigor for developing diagnostic tools.
Main Methods:
- Isolation of miRNAs from human plasma and serum samples.
- Application of different miRNA profiling techniques.
- Case examples from cardiovascular and pulmonary disease research.
Main Results:
- Key procedural considerations for successful miRNA isolation were identified.
- Advantages and disadvantages of various miRNA profiling methods were highlighted.
- The importance of methodological standardization was demonstrated.
Conclusions:
- Circulating miRNAs hold significant potential as diagnostic biomarkers.
- Strict adherence to methodological protocols is essential for reliable miRNA-based diagnostics.
- Further research focusing on standardization will advance the clinical application of miRNAs.
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