Abstract

Cardiac troponin T (cTnT) is considered a clinical standard for its high specificity and sensitivity when diagnosing acute myocardial infarction; however, most studies on the electrical sensors of cardiac troponin biomarkers have focused on cTnI rather than cTnT. This study presents label-free, low-cost, transparent, and flexible aptamer-based immunosensors for the electrical detection of cTnT using reduced graphene oxide (rGO) sheets. GO was first deposited by AC dielectrophoresis between two predefined source and drain electrodes on a 3-aminopropyltriethoxylsilane-modified polyethylene terephthalate substrate. The GO was then reduced using hydrazine vapour without damaging the substrate, resulting in uniform, controlled, and stable deposition of rGO sheets, and demonstrating more stability than those directly deposited by dielectrophoresis. Amine-modified single-strand DNA aptamers against cTnT were immobilized onto the rGO channels. The relative resistance change of this sensor owing to the attachment of cTnT was quantified as the cTnT concentration decreased from 10 ng/mL to 1 pg/mL in phosphate buffered saline (PBS) and 10-fold diluted human serum in PBS, with the limits of detection being 1.2 pg/mL and 1.7 pg/mL, respectively, which is sufficiently sensitive for clinical applications. High-yield and rapid fabrication of the present rGO sensors will have significant influences on scaled-up fabrication of graphene-based sensors.

Highlights

  • Cardiovascular disease is one of leading causes of mortality and morbidity globally

  • Raman spectra of the graphene oxide (GO) and reduced graphene oxide (rGO) sheets between Cr/Au electrodes are shown in Fig. S1A,B, respectively

  • We have demonstrated a low-cost, flexible, highly sensitive and label-free electrical aptasensors using DEP-deposited GO and its subsequent reduction to detect the cardiac biomarker troponin T

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Summary

Introduction

Cardiovascular disease is one of leading causes of mortality and morbidity globally. The statistics based on World Health Organization (WHO) showed that 17.9 million deaths were attributed to this disease in 20151, with 7.3 million being due to acute myocardial infarction (AMI)[2]. Among several biomarkers for the detection of AMI, both cardiac troponins (I & T) are considered the “gold standard” owing to their high sensitivity and specificity for cardiac muscle damage[3]. CTnT values provide an accurate diagnosis of absolute infarct size in AMI4. After the onset of AMI, the concentrations of the cardiac troponins begin to rise within 4–6 h, elevated up to more than two weeks for cTnT and more than 5–7 days for cTnI5. Despite the widespread use of cTnT and cTnI as diagnostic tools for AMI, commercially available cTnI immunoassay kits show large variations (at least a 5-fold difference) in the measured concentrations among them[7]. Conventional immunoassays employed for the detection of cTnT include enzyme-linked immunosorbent assay[10], fluoroimmunoassay[11], radioimmunoassay[12], immunochromatographic tests[13], and www.nature.com/scientificreports/

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