Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs

摘要

class="kwd-title">Abbreviations: PCR, polymerase chain reaction; CABG, coronary artery bypass grafting; EDTA, ethylenediaminetetraacetic acid; hsa-miR-1-3p, 3p strand of mature Homo sapiens microRNA-1; hsa-miR-208a-3p, 3p strand of mature Homo sapiens microRNA-208a; RT-qPCR, reverse transcription quantitative real-time PCR; RT, reverse transcription; Cq, quantification cycle; TP, time point; tRNA, transfer RNA; cel-miR-39-3p, 3p strand of mature Caenorhabditis elegans microRNA-39; RNase, ribonuclease; RIN, RNA integrity number; qPCR, quantitative real-time PCR class="kwd-title">Keywords: Heparinized plasma, microRNA quantification, RT-qPCR efficiency, Biomarkers class="head no_bottom_margin" id="abs0015title">AbstractBackgroundmicroRNAs have recently been identified as powerful biomarkers of human disease. Reliable polymerase chain reaction (PCR)-based quantification of nucleic acids in clinical samples contaminated with polymerase inhibitor heparin requires deheparinization. However, the effects of deheparinization procedure on quantification of nucleic acids remain largely unknown. The aim of this study was to determine whether the deheparinization procedure completely eliminates the inhibition of amplification, while maintaining RNA integrity and technical variability of the measured microRNA levels.