Patent ID: 7105319

Claim:
A method for amplifying pab gene, a gene element of Mycobacterium tuberculosis , which employs an RNA amplification process, comprising the steps of: forming a cDNA with an RNA-dependent DNA polymerase using a specific sequence of an RNA derived from said pab gene present in a sample as a template, with a first primer having a sequence homologous to said specific sequence and a second primer having a sequence complementary to said specific sequence, wherein either the first or second primer has a sequence having the RNA polymerase promoter sequence added at its 5′-region, thereby producing an RNA-DNA double-strand; digesting the RNA of said RNA-DNA double-strand with Ribonuclease H to form a single-stranded DNA; and then forming a double-stranded DNA that includes the promoter sequence allowing transcription of said specific sequence or an RNA comprising a sequence complementary to said specific sequence with a DNA-dependent DNA polymerase using said single-stranded DNA as a template, said double-stranded DNA produces an RNA transcription product in the presence of an RNA polymerase, and said RNA transcription product is subsequently used as the template for the cDNA synthesis with said RNA-dependent DNA polymerase; said method is characterized in that an oligonucleotide comprising at least 10 contiguous bases of SEQ ID NO: 33 having a sequence homologous to a portion of the RNA sequence of the Mycobacterium tuberculosis pab gene to be amplified is used as the first primer and an oligonucleotide comprising at least 10 contiguous bases of SEQ ID NO: 40 having a sequence complementary to a portion of the RNA sequence of the Mycobacterium tuberculosis pab gene is used as the second primer.