Chip-based digital PCR as a novel detection method for quantifying microRNAs in acute myocardial infarction patients  

作　　者：Samuel ROBINSON Marie FOLLO David HAENE Maximilian MAULER Daniela STALLMANN Lukas Andreas HEGER Thomas HELBING Daniel DUERSCHMIED Karlheinz PETER Christoph BODE Ingo AHRENS Marcus HORTMANN 

机构地区：[1]Department of Cardiology and Angiology I, Heart Center Freiburg University, Faculty of Medicine, University of Freiburg, Germany [2]Department of Medicine, Monash University, Melbourne, Australia [3]Department of Medicine I, Lighthouse Core Facility, Medical Center- University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany [4]Faculty of Biology, University of Freiburg [5]Baker IDI Heart and Diabetes Institute, Melbourne, Australia [6]Augustinerinnen Hospital, Academic Teaching Hospital University of Cologne, Cologne, Germany

出　　处：《Acta Pharmacologica Sinica》2018年第7期1217-1227,共11页中国药理学报（英文版）

摘　　要：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/pL vs 1.1 copies/pL） 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/pL vs 59 copies/pL; P=O.0300）, whereas qRT-PCR was unable to reach significance （136.4 copies/pL vs 122.8 copies/pL; 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/IJL vs 8.5 copies/pL, P=0.0011; 0 copies/pL vs 19.4 copies/pL; P〈OoO001）. 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 fmiRNAs 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/pL vs 1.1 copies/pL） 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/pL vs 59 copies/pL; P=O.0300）, whereas qRT-PCR was unable to reach significance （136.4 copies/pL vs 122.8 copies/pL; 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/IJL vs 8.5 copies/pL, P=0.0011; 0 copies/pL vs 19.4 copies/pL; P〈OoO001）. 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 f

关 键 词：ST-segment elevation myocardial infarction ichaemia-reperfusion injury micro-RNAs chip-based digital PCR QRT-PCR 

分 类 号：Q78[生物学—分子生物学] TN304.23[电子电信—物理电子学]