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:
CreatorsEmailORCIDORCID Put Code
Robinson, S.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Follo, M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Haenel, D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mauler, M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Stallmann, D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Tewari, M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Duerschmied, D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Peter, K.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Bode, C.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ahrens, I.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hortmann, M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
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:
KeywordsLanguage
PERCUTANEOUS CORONARY INTERVENTION; CIRCULATING MICRORNAS; CARDIOVASCULAR-DISEASE; PROGNOSTIC VALUE; EARLY-DIAGNOSIS; TROPONIN-T; SIZE; ELEVATION; BIOMARKER; INJURYMultiple languages
Cardiac & Cardiovascular SystemsMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/18928

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