Patent ID: 7049067

Claim:
A method for amplifying an RNA derived from HIV-1, which comprises the steps: (A) incubating RNA of a sample suspected to contain said RNA in the presence of an RNA-dependent DNA polymerase under conditions sufficient to synthesize a cDNA-RNA molecule therefrom, wherein said incubation is conducted in the presence of a first primer containing a sequence complementary to a specific sequence of said RNA and a second primer containing a sequence homologous to the specific sequence either of which additionally has a promoter sequence for the RNA polymerase at the 5′ end, (B) incubating said cDNA-RNA molecule in the presence of a ribonuclease H activity under conditions sufficient to degrade the RNA of said molecule thereby forming a single-stranded DNA molecule, (C) forming a double-stranded DNA molecule having a promoter sequence which can be transcribed into an RNA molecule comprising said specific base sequence or a sequence complementary to said specific base sequence by incubating said single-stranded DNA molecule in the presence of a DNA-dependent DNA polymerase, and (D) incubating said formed double-stranded DNA molecule in the presence of an RNA polymerase under conditions sufficient to produce an RNA transcript, wherein said RNA transcript acts as a template in a subsequent cDNA synthesis mediated by an RNA-dependent DNA polymerase, wherein said first primer is an oligonucleotide comprising at least 10 consecutive bases in any of SEQ ID NOS: 14 or 17, and the second primer is an oligonucleotide comprising at least 10 consecutive bases of SEQ ID NO: 27, to thereby amplify said RNA derived from HIV-1.