Universität zu Köln

Robinson, S., Follo, M., Haenel, D., Mauler, M., Stallmann, D., Tewari, M., Duerschmied, D., Peter, K., Bode, C., Ahrens, I. and Hortmann, M. (2018). Droplet digital PCR as a novel detection method for quantifying microRNAs in acute myocardial infarction. Int. J. Cardiol., 257. S. 247 - 255. CLARE: ELSEVIER IRELAND LTD. ISSN 1874-1754

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Abstract

Background: micro-RNAs have shown promise as potential biomarkers for acute myocardial infarction and ischemia-reperfusion injury (I/R). Most recently droplet digital polymerase chain reaction (ddPCR) has been introduced as a more reliable and reproducible method for detecting micro-RNAs. Aims: We aimed to demonstrate the improved technical performance and diagnostic potential of ddPCR by measuring micro-RNAs in ST-elevation myocardial infarction (STEMI). Methods: A dilution series was performed in duplicate on synthetic Caenorrhabditis elegans-miR-39, comparing quantitative real-time PCR (qRT-PCR) and ddPCR. We used ddPCR and qRT-PCR to quantify the serum levels of miR-21, miR-208a and miR-499 between STEMI patients (n=24) and stable coronary artery disease (CAD) patients (n = 20). In STEMI, I/R injury was assessed via measurement of ST-segment resolution. Results: In the dilution series, ddPCR demonstrated superior coefficient of variation (12.1% vs. 32.9%) and limit of detection (0.9325 vs. 2.425copies/mu l). In the patient cohort, ddPCR demonstrated greater differences inmiR-21 levels (2190.5 vs. 484.7 copies/mu l; p = 0.0004 for ddPCR and 136.4 vs. 122.8 copies/mu l; p = 0.2273 for qRT-PCR) and in miR-208a (0 vs. 24.1 copies/mu l, p = 0.0013 for ddPCR and 0 vs. 0 copies/mu l, p = 0.0032 for qRT-PCR), with similar differences observed in miR-499 levels (9.4 vs. 81.5 copies/mu l, p < 0.0001 for ddPCR and 0 vs. 19.41 copies/mu l, p < 0.0001 for qRT-PCR). ddPCR also more accurately defined STEMI for all miRNAs (area under the curve (AUC) of 0.8021/0.7740/0.9063 for miR-21/208a/499 with ddPCR vs. AUC of 0.6083/0.6917/0.8417 with qRTPCR). However, there was no association between miR-21/208a/499 levels and ischemia-reperfusion injury. Conclusion: ddPCR demonstrates superiority in both technical performance and diagnostic potential compared to qRT-PCR. Ultimately, this supports its use as a diagnosticmethod for quantifyingmicro-RNAs, particularly in large multi-center trials. (C) 2017 Elsevier B.V. All rights reserved.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Robinson, S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Follo, M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Haenel, D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Mauler, M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Stallmann, D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Tewari, M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Duerschmied, D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Peter, K. UNSPECIFIED UNSPECIFIED UNSPECIFIED Bode, C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Ahrens, I. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hortmann, M. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-189285
DOI:	10.1016/j.ijcard.2017.10.111
Journal or Publication Title:	Int. J. Cardiol.
Volume:	257
Page Range:	S. 247 - 255
Date:	2018
Publisher:	ELSEVIER IRELAND LTD
Place of Publication:	CLARE
ISSN:	1874-1754
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PERCUTANEOUS CORONARY INTERVENTION; CIRCULATING MICRORNAS; CARDIOVASCULAR-DISEASE; PROGNOSTIC VALUE; EARLY-DIAGNOSIS; TROPONIN-T; SIZE; ELEVATION; BIOMARKER; INJURY Multiple languages Cardiac & Cardiovascular Systems Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18928