Universität zu Köln

Robinson, Samuel, Follo, Marie, Haenel, David, Mauler, Maximilian, Stallmann, Daniela, Andreas, Lukas Heger, Helbing, Thomas, Duerschmied, Daniel ORCID: 0000-0001-5249-4012, Peter, Karlheinz, Bode, Christoph, Ahrens, Ingo and Hortmann, Marcus (2018). Chip-based digital PCR as a novel detection method for quantifying microRNAs in acute myocardial infarction patients. Acta Pharmacol. Sin., 39 (7). S. 1217 - 1228. SHANGHAI: ACTA PHARMACOLOGICA SINICA. ISSN 1745-7254

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Abstract

miRNAs have shown promise as potential biomarkers for acute myocardial infarction (AMI). However, the current used quantitative real-time PCR (qRT-PCR) allows solely for relative expression of nucleic acids and it is susceptible to day-to-day variability, which has limited the validity of using the miRNAs as biomarkers. In this study we explored the technical qualities and diagnostic potential of a new technique, chip-based digital PCR, in quantifying the miRNAs in patients with AMI and ischaemia-reperfusion injury (I/R). In a dilution series of synthetic C elegans-miR-39, chip-based digital PCR displayed a lower coefficient of variation (8.9% vs 46.3%) and a lower limit of detection (0.2 copies/mu L vs 1.1 copies/mu L) compared with qRT-PCR. In the serum collected from 24 patients with ST-elevation myocardial infarction (STEMI) and 20 patients with stable coronary artery disease (CAD) patients after percutaneous coronary intervention (PCI), we used qRT-PCR and multiplexed chip-based digital PCR to quantify the serum levels of miRNA-21 and miRNA-499 as they have been validated in AMI in prior studies. In STEMI, I/R injury was assessed via measurement of ST-segment resolution (ST-R). Chip-based digital PCR revealed a statistical significance in the difference of miR-21 levels between stable CAD and STEMI groups (118.8 copies/mu L vs 59 copies/mu L; P=0.0300), whereas qRT-PCR was unable to reach significance (136.4 copies/mu L vs 122.8 copies/mu L; P=0.2273). For miR-499 levels, both chip-based digital PCR and qRT-PCR revealed statistically significant differences between stable CAD and STEMI groups (2 copies/mu L vs 8.5 copies/mu L, P=0.0011; 0 copies/mu L vs 19.4 copies/mu L; P<0.0001). There was no association between miR-21/499 levels and ST-R post-PCI. Our results show that the chip-based digital PCR exhibits superior technical qualities and promises to be a superior method for quantifying miRNA levels in the circulation, which may become a more accurate and reproducible method for directly quantifying miRNAs, particularly for use in large multi-centre clinical trials.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Robinson, Samuel UNSPECIFIED UNSPECIFIED UNSPECIFIED Follo, Marie UNSPECIFIED UNSPECIFIED UNSPECIFIED Haenel, David UNSPECIFIED UNSPECIFIED UNSPECIFIED Mauler, Maximilian UNSPECIFIED UNSPECIFIED UNSPECIFIED Stallmann, Daniela UNSPECIFIED UNSPECIFIED UNSPECIFIED Andreas, Lukas Heger UNSPECIFIED UNSPECIFIED UNSPECIFIED Helbing, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Duerschmied, Daniel UNSPECIFIED orcid.org/0000-0001-5249-4012 UNSPECIFIED Peter, Karlheinz UNSPECIFIED UNSPECIFIED UNSPECIFIED Bode, Christoph UNSPECIFIED UNSPECIFIED UNSPECIFIED Ahrens, Ingo UNSPECIFIED UNSPECIFIED UNSPECIFIED Hortmann, Marcus UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-181421
DOI:	10.1038/aps.2017.136
Journal or Publication Title:	Acta Pharmacol. Sin.
Volume:	39
Number:	7
Page Range:	S. 1217 - 1228
Date:	2018
Publisher:	ACTA PHARMACOLOGICA SINICA
Place of Publication:	SHANGHAI
ISSN:	1745-7254
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ST-SEGMENT ELEVATION; CREATINE KINASE-MB; CIRCULATING MICRORNAS; QUANTIFICATION; QUANTITATION; EXPRESSION; BIOMARKERS; RESOLUTION; EFFICACY Multiple languages Chemistry, Multidisciplinary; Pharmacology & Pharmacy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18142