The invention discloses a real-
time reaction monitoring method and a real-
time reaction monitoring system for
DNA (
Deoxyribose Nucleic Acid) base removal sites and
small molecule toxic compounds, and belongs to the technical field of biological analysis. The method comprises the following steps: constructing a compound formed by combining a
uracil-containing
DNA probe and an electrochemical active marker, and hydrolyzing
uracil by using UDG
enzyme to form a base removal site. A
nanopore single-molecule detection technology is adopted, a compound is driven by
voltage to pass through a hole, and a characteristic current
signal is generated to establish a baseline map.
Hydroxylamine derivatives are introduced to react with base removal sites, and dynamic interaction is analyzed according to
time domain change of current signals. The
system comprises a labeled compound preparation module, an enzymatic reaction module, a
nanopore detection module, a
small molecule compound reaction module and a central control and
data processing module, all the modules cooperate, full-process
automation from probe preparation to dynamic reaction monitoring is achieved, traditional sensitivity limitation is broken through, and a high-precision detection means is provided for studying the
DNA damage and toxic compound action mechanism.