Method for detecting target rna by utilizing nicking/extension chain reaction system-based isothermal nucleic acid amplification

Abstract

The present invention relates to a detection method for detecting a target RNA contained in a sample with high sensitivity by using nicking/extension chain reaction system-based isothermal nucleic acid amplification (NESBA) that uses activity of a cleavage enzyme and a DNA polymerase. The NESBA of the present invention is a new concept isothermal target RNA detection method that realizes higher amplification efficiency than the existing NASBA technology and is deemed to be utilizable as a new concept diagnosis technology that can replace conventional target RNA detection technologies.

What is claimed is: 
     
         1 . A composition for detecting a target RNA, the composition comprising:
 (i) a first primer comprising a sequence complementary to the target RNA, a T7 promoter sequence, and a DNA nicking enzyme recognition nucleotide sequence; and   (ii) a second primer comprising a sequence complementary to DNA complementary to the target nucleic acid and a DNA nicking enzyme recognition nucleotide sequence.   
     
     
         2 . The composition according to  claim 1 , further comprising a reverse transcriptase, T7 RNA polymerase, a DNA endonuclease, DNA polymerase, and dNTP. 
     
     
         3 . A kit for detecting a target RNA, the kit comprising:
 (i) a first primer comprising a sequence complementary to the target RNA, a T7 promoter sequence, and a DNA nicking enzyme recognition nucleotide sequence; and   (ii) a second primer comprising a sequence complementary to DNA complementary to the target nucleic acid and a DNA nicking enzyme recognition nucleotide sequence.   
     
     
         4 . The kit according to  claim 3 , further comprising a reverse transcriptase, T7 RNA polymerase, a DNA endonuclease, DNA polymerase, and dNTP.