Patent ID: 8263334

Claim:
A method of nucleic acid sequence detection for detecting the presence of a particular target nucleic acid sequence to be detected in a sample, the method comprising: a design step of designing first and second oligonucleotide strands in advance such that the second oligonucleotide strand has a complementary nucleic acid sequence to an elongation reaction product dissociated from a duplex formed through elongation reaction using the first oligonucleotide strand as a primer and the target nucleic acid sequence as a template after complementary binding of the target nucleic acid sequence and the first oligonucleotide strand; an immobilization step of anchoring the first and second oligonucleotide strands onto a substrate by immobilizing the 5′-ends thereof onto the substrate; a first elongation reaction step of complementarily binding the first oligonucleotide strand to the target nucleic acid sequence and performing first elongation reaction using the first oligonucleotide strand as a primer and the target nucleic acid sequence as a template; a first dissociation step of dissociating, from the target nucleic acid sequence, the first elongation reaction product extended in the first elongation reaction step; a first cross-linking step of binding the dissociated first elongation reaction product to the second oligonucleotide strand to form a cross-linked structure between the first and second oligonucleotide strands; a second elongation reaction step of performing second elongation reaction using the second oligonucleotide strand as a primer and the first elongation reaction product as a template; a second dissociation step of dissociating therefrom the second elongation reaction product extended in the second elongation reaction step; a second cross-linking step of performing the first elongation reaction step to the second dissociation step on a plurality of the first and second oligonucleotide strands and binding a first oligonucleotide strand different from the first oligonucleotide strand to the dissociated second elongation reaction product to form a cross-linked structure between the second and first oligonucleotide strands; a repetitive cross-linking step of repeating the first and second cross-linking steps between a plurality of the first and second oligonucleotide strands; a reticulated space formation step of forming a reticulated space by the repetitive cross-linking step, wherein a plurality of the cross-linked structures are intertwined with each other; a capture step of capturing a ligand within the reticulated space through physical adsorption; a coloring step of causing the captured ligand to color with an active substance reactive to the ligand; and a detection step of detecting a color signal derived from the coloring to detect the target nucleic acid sequence.