A method of detecting a specific target polynucleotide in an isothermal amplification reaction coupled with reverse transcription, the method comprising: (a) contacting a target polynucleotide molecule in a sample with a reverse transcriptase primer in the presence of a reverse transcriptase and dNTPs under conditions permissive for cDNA synthesis, thereby generating a cDNA; (b) contacting the cDNA with (i) forward and reverse primers, wherein each primer comprises a 5'-terminal region carrying at least one nicking enzyme recognition sequence, a 3'-terminal region which specifically binds the cDNA in the presence of a nicking enzyme, and one or more 2' modified nucleotides at the end of the 3'-terminal region, wherein the one or more 2' modified nucleotides is one or more of 2'-O-methyl, 2'-methoxyethoxy, 2'-fluoro, 2'-alkyl, 2'-allyl, 2'-O-[2-(methylamino)-2-oxoethyl], 2'-hydroxyl, 4'-CH 2 -O-2'-bridge, 4'-(CH 2 ) 2 -O-2' -bridge, 2'-LNA and 2'-O-(N-methylcarbamate), (ii) dNTPs, (iii) a detectable oligonucleotide probe, and (iv) a strand-displacement polymerase under conditions permissive for the isothermal amplification of the cDNA; and (c) detecting a signal specific for detectable oligonucleotide probe hybridization to the amplicon, wherein detection of the signal indicates the presence or quantity of the target polynucleotide present in the sample and failure to detect the signal indicates the absence of target polynucleotide in the sample. (a) contacting a target polynucleotide molecule in a sample with a reverse transcriptase primer in the presence of a reverse transcriptase and dNTPs under conditions permissive for cDNA synthesis, thereby generating a cDNA; (b) contacting the cDNA with (i) forward and reverse primers, wherein each primer comprises a 5'-terminal region carrying at least one nicking enzyme recognition sequence, a 3'-terminal region which specifically binds the cDNA in the presence of a nicking enzyme, and one or more 2' modified nucleotides at the end of the 3'-terminal region, wherein the one or more 2' modified nucleotides is one or more of 2'-O-methyl, 2'-methoxyethoxy, 2'-fluoro, 2'-alkyl, 2'-allyl, 2'-O-[2-(methylamino)-2-oxoethyl], 2'-hydroxyl, 4'-CH 2 -O-2'-bridge, 4'-(CH 2 ) 2 -O-2' -bridge, 2'-LNA and 2'-O-(N-methylcarbamate), (ii) dNTPs, (iii) a detectable oligonucleotide probe, and (iv) a strand-displacement polymerase under conditions permissive for the isothermal amplification of the cDNA; and (i) forward and reverse primers, wherein each primer comprises a 5'-terminal region carrying at least one nicking enzyme recognition sequence, a 3'-terminal region which specifically binds the cDNA in the presence of a nicking enzyme, and one or more 2' modified nucleotides at the end of the 3'-terminal region, wherein the one or more 2' modified nucleotides is one or more of 2'-O-methyl, 2'-methoxyethoxy, 2'-fluoro, 2'-alkyl, 2'-allyl, 2'-O-[2-(methylamino)-2-oxoethyl], 2'-hydroxyl, 4'-CH 2 -O-2'-bridge, 4'-(CH 2 ) 2 -O-2' -bridge, 2'-LNA and 2'-O-(N-methylcarbamate), (ii) dNTPs, (iii) a detectable oligonucleotide probe, and (iv) a strand-displacement polymerase (c) detecting a signal specific for detectable oligonucleotide probe hybridization to the amplicon, wherein detection of the signal indicates the presence or quantity of the target polynucleotide present in the sample and failure to detect the signal indicates the absence of target polynucleotide in the sample. <EOS>