Abstract
A novel aptamer-based enzyme-linked assay for the detection of adenosine triphosphate( ATP) was established based on the specific recognition of aptamer towards target and the efficient catalytic ability of horseradish peroxidase( HRP). Aptamer binding with targets caused that duplex DNA containing short chain DNA and aptamer dissociated and dissociated DNA was captured by another DNA immobilized on other plate through hybridization. A fluorescein isothiocyanate( FITC) was tagged on dissociated DNA,a HRP as signal transduction element was linked on microplate through specific interactions with anti-FITC-HRP and catalyzed methyenedianiline( TMB) substrate to produce color change. The detection of ATP was achieved by color change and absorbance change at 450 nm. This method has good selectivity for ATP recognition interfering from other proteins such as GTP,UTP and CTP and the detection can be carried in complex sample such as 10% and 50% serum. Experimental results show that there is a good linear relationship with ATP concentration ranged from 50 to 400 nmol / L,and the detection limit is 26 nmol / L.
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