The invention provides an ATP chemiluminiscence detection method based on an
enzyme digestion assisted
label-free
aptamer sensor, which comprises the following steps: immobilizing an
aptamer on the surface of a magnetic
microsphere, and specifically binding the
aptamer with
adenosine triphosphate, then, degrading the single-stranded
DNA specifically and gradually into a free basic group from the 3' end by utilizing
exonuclease T, and reducing a background
signal remarkably based on the
exonuclease, so that a quantitative relation between the quantity of the aptamer detected on the surface of the magnetic
microsphere and the concentration of the ATP to be detected is obtained; and finally, carrying out chemiluminiscence detection on the aptamer sensor. Based on the principle that
guanine base (G) on an ATP aptamer chain and a
chemiluminescence reagent benzoyl
formaldehyde (PG) are subjected to an instantaneous
derivatization reaction to generate
chemiluminescence, the marking link of other
chemiluminescence markers is omitted, the operation steps are simplified on the basis of ensuring the sensitivity, the detection time is shortened, and the detection efficiency is improved. In addition, the used chemiluminescence
analysis method is wide in
linear range, simple in equipment and easy to operate, so that full-automatic detection is expected to be realized.