Measuring method of cytokeratin 19mRNA

    公开(公告)号:JP5481841B2

    公开(公告)日:2014-04-23

    申请号:JP2008303637

    申请日:2008-11-28

    IPC分类号: C12Q1/68 C12N15/09

    摘要: Disclosed is a method of amplifying and detecting cytokeratin 19 mRNA in RNA amplification process, comprising: a step for forming a double-stranded DNA containing a promoter sequence with a reverse transcriptase by use of a combination of oligonucleotides consisting of a first primer having a sequence homologous to a portion of cytokeratin 19 mRNA and a second primer having a complementary sequence, wherein the promoter sequence is added to the 5'-end of either the first primer or the second primer, forming an RNA transcription product by use of an RNA polymerase with using the double-stranded DNA as template, and forming the double-stranded DNA by use of a reverse transcriptase by continuing to use the RNA transcription product as a template of DNA synthesis, by measuring an amount of amplified RNA produced over time with an oligonucleotide probe designed so that signal properties change with the formation of a complementary double strand with the amplified RNA.