Abstract
BackgroundIn this paper, a simple, enzyme-free, label-free fluorescence, high sensitivity logic gate hairpin aptasensor was developed for adenosine triphosphate (ATP) detection based on graphene oxide (GO) and PicoGreen dye.MethodsUsing single-strand deoxyribonucleic acid (DNA) and adenosine triphosphate (ATP) as input signal and fluorescence signal as output signal, if single-strand DNA (DNA-L), single-strand DNA (DNA-S), and ATP were present at the same time, one segment of DNA-L formed a hairpin ring with ATP, and the other segment of DNA-L formed a completely complementary hairpin stem with DNA-S. The hairpin DNA was detached from the GO surface, and PicoGreen dye was embedded into the hairpin stem, and the fluorescence signal was enhanced. The molecular logic gate was constructed through the establishment of logic histogram, logic circuit, truth table, and logic formula. The biosensor-related performances including sensitivity, selectivity, and linearity were investigated, respectively.ResultsWe have successfully constructed a AND logic gate. The detection limit of ATP is 138.0 pmol/L (3σ/slope) with detection range of 50–500 nmol/L (R2 = 0.98951), and its sensitivity is 4.748 × 106–6.875 × 108 a.u. (mol/L)−1.ConclusionsThe logic gate hairpin aptamer sensor has the advantages of high sensitivity, low detection limit, and low cost, and can be successfully applied to the detection of adenosine triphosphate (ATP) in actual human urine samples.
Highlights
Adenosine triphosphate (ATP) is associated with hypoglycemia, Parkinson’s disease, hypoxia, and ischemia (Aw and Jones 1985; Agardh et al 1978; Katsura et al 1993)
Single-stranded deoxyribonucleic acid (DNA) was adsorbed on the surface of graphene oxide (GO) through π-π stacking, fluorescence resonance energy transfer (FRET) occurred between PicoGreen dye combined with singlestranded DNA and GO, and fluorescence was quenched
The distance between PicoGreen embedded in double-stranded DNA and GO increased, and FRET could not occur between PicoGreen and GO
Summary
Adenosine triphosphate (ATP) is associated with hypoglycemia, Parkinson’s disease, hypoxia, and ischemia (Aw and Jones 1985; Agardh et al 1978; Katsura et al 1993). The study of ATP detection method has important clinical significance. The signal amplification and background signal suppression are two main strategies for improving DNA fluorescence sensors sensitivity. Common methods based on signal amplification can effectively increase sensor sensitivity, including rolling circle replication (RCR) (Zhou et al 2015), strand displacement amplification (SDA) (Chen et al 2016; Wang et al 2013; Wei et al 2016; Wang et al 2017), and DNA enzymes (Zeng et al 2015; Lu et al 2010; He et al 2017; Ning et al 2017), etc. A simple, enzyme-free, label-free fluorescence, high sensitivity logic gate hairpin aptasensor was developed for adenosine triphosphate (ATP) detection based on graphene oxide (GO) and PicoGreen dye
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