The invention relates to the technical field of
nucleic acid detection, in particular to a
nucleic acid target sequence detection method and
system based on
solid phase hybridization and
enzyme digestion recognition and application. The method comprises the following steps: preparing target
nucleic acid with a detectable marker; hybridizing with a specific probe fixed on a
solid-phase carrier; treating the
solid phase carrier with an
enzyme treatment system comprising a cleavage mismatched
endonuclease to specifically remove
hybrid double strands that are not completely matched with the probe; and finally, detecting a mark
signal reserved on the carrier so as to judge the state of the target sequence. According to the method, an
enzyme digestion verification step is introduced after traditional solid-phase hybridization, and non-specific hybridization is eliminated from a
molecular level by utilizing high recognition capability of enzyme on nucleic acid mismatch or structural
distortion, so that ultrahigh
specific detection close to single base resolution is realized. The technical platform is high in universality, and can be applicable to detection of various types of targets such as
single nucleotide polymorphism (SNP),
point mutation, short
tandem repeat (STR) and
pathogen genes only by replacing the probes. The method is simple and convenient to operate, can be easily integrated with isothermal amplification,
test strips and other simple carriers, is suitable for on-site
rapid detection, and has wide application prospects in the fields of molecular diagnosis, forensic
medicine identification,
pathogen monitoring and the like.