HOMOGENEOUS ISOTHERMAL AMPLIFICATION AND DETECTION OF NUCLEIC ACIDS USING A TEMPLATE SWITCH OLIGONUCLEOTIDE

摘要

An isothermal, transcription-based nucleic acid amplification method based on the formation of a target-dependent nucleic acid species by template switching. This product species can be amplified further to produce multiple copies of both double stranded DNA products and single stranded RNA product. The single stranded RNA amplification products are the same sense as the target sequence. Also provided is a branch migration inhibition based procedure for scanning nucleic acid sequences using the strand switch isothermal amplification. A template switch oligonucleotide used in the amplification includes a 3' region capable of hybridizing to a target sequence and a 5' region which does not hybridize to the target. The 5' region includes a promoter sequence of DNA dependent RNA polymerase. A first primer and a second primer may be used during amplification and detection. The first primer is capable of hybridizing to the target and the second primer is not target dependent. For detection, either the first primer or the second primer has a label.