Practical Procedures for Improving Detection of Circulating miRNAs in Cardiovascular Diseases

Chia Chun Chen; Chen Ching Peng; Pei Chun Fan; Pao Hsien Chu; Yu Sun Chang; Chih Hsiang Chang

doi:10.1007/s12265-020-10019-2

Practical Procedures for Improving Detection of Circulating miRNAs in Cardiovascular Diseases

Chia Chun Chen^*, Chen Ching Peng, Pei Chun Fan, Pao Hsien Chu, Yu Sun Chang, Chih Hsiang Chang

^*Corresponding author for this work

Molecular Medicine Research Center

Chang Gung University

Research output: Contribution to journal › Journal Article › peer-review

4 Scopus citations

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. [Figure not available: see fulltext.]

Original language	English
Pages (from-to)	977-987
Number of pages	11
Journal	Journal of Cardiovascular Translational Research
Volume	13
Issue number	6
DOIs	https://doi.org/10.1007/s12265-020-10019-2
State	Published - 12 2020

Bibliographical note

Publisher Copyright:
© 2020, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Hemolysis
Heparin
Heparinase
MicroRNA
Normalization

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/s12265-020-10019-2

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title = "Practical Procedures for Improving Detection of Circulating miRNAs in Cardiovascular Diseases",

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. [Figure not available: see fulltext.]",

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AU - Chen, Chia Chun

AU - Peng, Chen Ching

AU - Fan, Pei Chun

AU - Chu, Pao Hsien

AU - Chang, Yu Sun

AU - Chang, Chih Hsiang

PY - 2020/12

Y1 - 2020/12

N2 - 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. [Figure not available: see fulltext.]

AB - 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. [Figure not available: see fulltext.]

KW - Hemolysis

KW - Heparin

KW - Heparinase

KW - MicroRNA

KW - Normalization

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IS - 6

ER -

Practical Procedures for Improving Detection of Circulating miRNAs in Cardiovascular Diseases

Abstract

Bibliographical note

Keywords

UN SDGs

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