Nucleic acid detection method, detection probe, detection probe set, and nucleic acid quantification method

Abstract

A method for detecting a target nucleic acid, comprising: (a) contacting a nucleic acid sample comprising a target nucleic acid, comprising a first portion and a second portion, with: (i) a detection probe, wherein the detection probe is labeled with a labeling substance and comprises a nucleic acid sequence that forms a stem-loop structure and having a 5′ protruding end or a 3′ protruding end that is capable of hybridizing to the second portion, and (ii) a capture probe comprising a nucleic acid sequence capable of hybridizing to the first portion, wherein the capture probe is immobilized to a substrate, under conditions to form a target nucleic acid-detection probe-capture probe complex by hybridizing the second portion to the detection probe and hybridizing the first portion to the capture probe; (b) ligating a first end of the detection probe with an end of the target nucleic acid and ligating a second end of the detection probe with an end of the capture probe; and (c) detecting the labeling substance of the nucleic acid-detection probe-capture probe complex formed on the substrate.

What is claimed is: 
     
         1 . A probe set for detecting a target nucleic acid comprising a first portion and a second portion, comprising:
 a detection probe comprising a first stem forming portion, a second stem forming portion, a loop portion, and a 5′ protruding end or 3′ protruding end that comprises a sequence capable of hybridizing to the second portion of the target nucleic acid, wherein the first and second stem forming portions are complementary to each other, the loop portion is located between the first and second stem forming portions, the loop portion is labeled with a labeling substance; and   a capture probe comprising a sequence that hybridizes to the first portion of the target nucleic acid, wherein the capture probe is immobilized to a substrate.   
     
     
         2 . The probe set according to  claim 1 , wherein the target nucleic acid is miRNA. 
     
     
         3 . The probe set according to  claim 1 , wherein the capture probe comprises a spacer at one end that binds to the substrate.