Patent ID: 6756198
Filing Date: 2004-06-29
Classification: C12Q

Abstract:
An amplification step which comprises:forming an RNA-DNA duplex by forming a cDNA by the action of an RNA-dependent DNA polymerase using a specific sequence of RNA from Hepatitis C virus in a sample as a template, a first primer having a sequence homologous to the specific sequence, and a second primer having a sequence complementary to the specific sequence (wherein either primer of the first or second primers has a sequence to which a promoter sequence of an RNA polymerase is added at the 5â€² end), forming a single-stranded DNA by degrading the RNA-DNA duplex by ribonuclease H, forming a double-stranded DNA having a promoter sequence for transcribing an RNA comprising the above RNA sequence or a sequence complementary to the above RNA sequence by a DNA-dependent DNA polymerase using the single-stranded DNA as a template, and forming an RNA transcription product from the double-strand DNA in the presence of an RNA polymerase, the RNA transcription product successively acting as a template for the formation of the single-stranded DNA by the above RNA-dependent DNA polymerase, wherein the oligonucleotide of SEQ ID NO: 11 is used as the first primer and the oligonucleotide of SEQ ID NO: 6 or 7 as the second primer; the oligonucleotide of SEQ ID NO: 12 is used as the first primer and the oligonucleotide of SEQ ID NO: 7 as the second primer; or the oligonucleotide of SEQ ID NO: 13 is used as the first primer and the oligonucleotide of SEQ ID NO: 9 as the second primer.