Urine is the most appropriate body fluid for analysis because it is easily and less-invasively obtained than blood; thus, urinary miRNAs can better represent the local stage of the disease and might grow up to be a new class of noninvasive biomarkers of postmyocardial infarction (MI). Monofunctionalized Au nanoparticles (AuNPs) with only one selective DNA at a specific location are more promising in nanotechnology. This study developed a urinary miRNA ultratrace detection strategy based on single-target DNA-functionalized AuNPs for the noninvasive prognosis of post-MI. The AuNPs were designed with only single-stranded biotinylated DNA complementary to the target miRNA through a ratio-optimized stoichiometric method for the first time. Combined with the duplex specific nuclease-assisted target recycling amplification, the single-target DNA-functionalized AuNPs for the first time were used in inductively coupled plasma-mass spectrometry for the determination of urinary miRNA with high sensitivity. After optimizing the reaction conditions, a linear detection range between 1 fM and 10 pM for miR-155 and a detection limit of 0.47 fM were obtained. Finally, the target miR-155 in urine samples collected from MI rats was quantified and the level of miR-155 in MI groups was 30 times higher than in the control groups. The results suggest that urinary miR-155 could be a novel biomarker for the noninvasive diagnosis of MI.
Keywords: inductively coupled plasma−mass spectrometry; noninvasive prognosis; post-MI; single-target DNA-functionalized AuNPs; urinary miR-155.