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Updated: Dec 20, 2025
Digital PCR for Quantifying Circulating MicroRNAs in Acute Myocardial Infarction and Cardiovascular Disease
Published on: July 3, 2018
Practical Procedures for Improving Detection of Circulating miRNAs in Cardiovascular Diseases
Chia-Chun Chen1,2, Chen-Ching Peng3, Pei-Chun Fan4,5
1Molecular Medicine Research Center, Chang Gung University, No. 259 Wen-Hwa 1st Road, Kwei-Shan, Taoyuan, 33302, Taiwan. chenchiachun@mail.cgu.edu.tw.
Abstract:
Hemolysis has been known to affect the measurement of circulating biomarkers. In this study, clinically applicable procedures for microRNA (miRNA) detection in serum samples of acute myocardial infarction patients were established. The 89 samples from patients admitted to the coronary care unit were collected. These samples obtained from heparin-treated and untreated patients were subjected to heparinase digestion prior to miRNA measurements by multiplex RT-qPCR. The good reproducibility of miRNA detection after heparinase digestion (average R2 = 0.97) indicated that this method can be used routinely for samples regardless of heparin medication. Additionally, the degree of hemolysis in these samples was highly related to the hemoglobin absorbance at 414 nm. Based on the hemoglobin absorbance, five hemolysis-associated miRNAs were identified in our data normalized with respect to both the spike-in control and the RNA amount in a given sample. Using these calibration procedures, miRNAs can be accurately quantified and identified for clinical samples. Graphical Abstract The practical procedures for miRNA detection in serum samples from the coronary care unit were established, and five hemolysis-associated miRNAs were accurately clarified through serial normalization.
Insights
This study establishes reliable microRNA (miRNA) detection in patient serum, even with hemolysis. Heparinase digestion and normalization methods ensure accurate quantification of hemolysis-associated miRNAs for clinical use.
Area of Science:
- Biochemistry
- Molecular Biology
- Clinical Diagnostics
Background:
- Hemolysis interferes with circulating biomarker measurements.
- Accurate microRNA (miRNA) detection is crucial for diagnosing conditions like acute myocardial infarction.
- Standardized protocols are needed for reliable miRNA analysis in clinical samples.
Purpose of the Study:
- To establish clinically applicable procedures for microRNA detection in serum samples from acute myocardial infarction patients.
- To address the impact of hemolysis and heparin on miRNA measurements.
- To identify hemolysis-associated miRNAs for improved diagnostic accuracy.
Main Methods:
- Collected 89 serum samples from coronary care unit patients.
- Applied heparinase digestion to samples from both heparin-treated and untreated patients.
- Performed microRNA measurements using multiplex RT-qPCR.
- Quantified hemolysis using hemoglobin absorbance at 414 nm.
- Normalized miRNA data using spike-in controls and RNA amount.
Main Results:
- Established reproducible miRNA detection after heparinase digestion (average R² = 0.97).
- Demonstrated that heparinase digestion allows routine miRNA measurement regardless of heparin medication.
- Identified a strong correlation between hemolysis degree and hemoglobin absorbance.
- Identified five hemolysis-associated miRNAs through normalization procedures.
Conclusions:
- Developed a robust method for routine miRNA quantification in clinical serum samples.
- Accurate identification of hemolysis-associated miRNAs is possible using the described calibration procedures.
- The findings support the clinical utility of miRNA analysis in acute myocardial infarction diagnostics, even with sample hemolysis.
