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    Detection of nucleic acid amplification
    2.
    发明授权
    Detection of nucleic acid amplification 有权
    检测核酸扩增

    公开(公告)号：US06258546B1

    公开(公告)日：2001-07-10

    申请号：US09602996

    申请日：2000-06-23

    申请人： Ray A. McMillian ,  Karen Eckert ,  Donald W. Copertino ,  Tobin J. Hellyer

    发明人： Ray A. McMillian ,  Karen Eckert ,  Donald W. Copertino ,  Tobin J. Hellyer

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6818 ,  C12Q1/6844 ,  C12Q2531/119 ,  C12Q2525/301 ,  C12Q2521/301 ,  C12Q2565/1025

    摘要： For use in nucleic acid amplification reactions, the detector oligonucleotides of the invention comprise a target binding sequence which is at least partially the same as the target binding sequence of an amplification primer present in the target amplification reaction, so that the detector oligonucleotide and the amplification primer compete for hybridization to the same sequence in the target. Hybridization of the amplification primer to the target upstream from the detector oligonucleotide generates a nickable restriction endonuclease recognition site. When this site is nicked and strand displacement occurs from the nick, both the 3′ end of the amplification primer and the detector oligonucleotide are displaced. The displaced detector oligonucleotide may then be detected as an indication of the presence of the target sequence, for example by unfolding of a fluorescently labeled secondary structure present in the detector oligonucleotide to reduce fluorescence quenching.

    摘要翻译： 为了用于核酸扩增反应，本发明的检测寡核苷酸包含与目标扩增反应中存在的扩增引物的靶结合序列至少部分相同的靶结合序列，使得检测寡核苷酸和扩增 引物竞争与目标中相同序列的杂交。 扩增引物与检测寡核苷酸上游的靶的杂交产生可切口的限制性内切核酸酶识别位点。 当该位点被切口并且从切口发生链置换时，扩增引物的3'末端和检测器寡核苷酸都被置换。 然后可以将位移的检测器寡核苷酸作为目标序列存在的指示进行检测，例如通过解折叠存在于检测器寡核苷酸中的荧光标记的二级结构以减少荧光猝灭。