In
the deoxyribonucleic acid (DNA) repair pathways, DNA repair
enzymes have great significance for genomic integrity. As one important
initiator of the base-excision repair pathway, the aberrant activity
of uracil-DNA glycosylase (UDG) is closely associated with many diseases.
Herein, we developed a simple distance-based device for visual detection
of UDG activity using a load-free DNA hydrogel. The DNA hydrogel consists
of polyacrylamide–DNA chains being bridged by a single-stranded
DNA crosslinker containing a responsive uracil base site. UDG can
recognize and remove the uracil, resulting in the cleavage effect
of the DNA crosslinker strand with the assistance of endonuclease
IV (Endo IV). Plugging one end of the capillary tube, the DNA hydrogel
acting as a filter membrane separator would control molecules to flow
into the tube. The integrity of the DNA hydrogel networks is affected
by the excision of UDG. Therefore, taking full advantage of membrane
filtration of the DNA hydrogel, the activity of UDG can be quantitatively
detected via reading the distance of the red indicator solution in
the capillary tube. Without any instruments and complicated procedures,
this method realizes high sensitivity and specificity for the detection
of UDG as low as 0.02 mU/mL and can even measure UDG in complex cell
samples. Additionally, this method is simple, universal, and can be
used to screen inhibitors, which shows great potential for point-of-care
testing, clinical diagnosis, and drug discovery.