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公开(公告)号：EP2351851A1
公开(公告)日：2011-08-03
申请号：EP09829212.1
申请日：2009-11-30
申请人： Tosoh Corporation
CPC分类号： C12Q1/6853 , C12Q1/6865 , C12Q2525/143 , C12Q2545/114 , C12Q2561/113 , C12Q2531/143 , C12Q2521/107
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.
摘要翻译： 本发明公开了在RNA扩增过程中扩增和检测细胞角蛋白19mRNA的方法，其包括：用逆转录酶形成含有启动子序列的双链DNA的步骤，其中使用由具有序列的第一引物 与细胞角蛋白19 mRNA的一部分同源，以及具有互补序列的第二引物，其中启动子序列被添加到第一引物或第二引物的5'末端，通过使用RNA聚合酶形成RNA转录产物 以双链DNA为模板，通过继续使用RNA转录产物作为DNA合成的模板，通过使用逆转录酶形成双链DNA，通过测量随时间产生的扩增RNA的量 寡核苷酸探针，其设计使得信号特性随着与扩增的RNA形成互补双链而改变。
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公开(公告)号：EP2351851B1
公开(公告)日：2015-07-08
申请号：EP09829212.1
申请日：2009-11-30
申请人： Tosoh Corporation
CPC分类号： C12Q1/6853 , C12Q1/6865 , C12Q2525/143 , C12Q2545/114 , C12Q2561/113 , C12Q2531/143 , C12Q2521/107
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公开(公告)号：EP2345741B1
公开(公告)日：2014-09-10
申请号：EP09819288.3
申请日：2009-10-06
申请人： Tosoh Corporation
CPC分类号： C12Q1/6886 , C12Q1/6851 , C12Q1/6865 , C12Q2600/158 , C12Q2525/143 , C12Q2563/173
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公开(公告)号：EP2345741A1
公开(公告)日：2011-07-20
申请号：EP09819288.3
申请日：2009-10-06
申请人： Tosoh Corporation
CPC分类号： C12Q1/6886 , C12Q1/6851 , C12Q1/6865 , C12Q2600/158 , C12Q2525/143 , C12Q2563/173
摘要： Disclosed is a method of amplifying and detecting mRNA of survivin gene in an RNA amplification process comprising: a step for forming a double-stranded DNA containing a promoter sequence by use of a combination of oligonucleotides consisting of a first primer having a sequence homologous with a portion of survivin 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 with a reverse transcriptase, 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, 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.
摘要翻译： 公开了在RNA扩增方法中扩增和检测存活蛋白基因的mRNA的方法，包括：通过使用由具有与第一引物序列同源的第一引物组成的寡核苷酸组合形成含有启动子序列的双链DNA的步骤 存在蛋白mRNA的部分和具有互补序列的第二引物，其中所述启动子序列用逆转录酶加入到第一引物或第二引物的5'-末端，通过使用RNA聚合酶形成RNA转录产物 使用双链DNA作为模板，并通过继续使用RNA转录产物作为DNA合成的模板，通过使用逆转录酶形成双链DNA，测量随着时间的推移用寡核苷酸产生的扩增的RNA的量 探针的设计使得信号特性随着扩增的RNA形成互补双链而发生变化。
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