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    • 3. 发明申请
    • PROBES FOR DETECTING POLYNUCLEOTIDES AND DETECTION METHOD
    • 用于检测聚合物和检测方法的探针
    • WO1998013524A1
    • 1998-04-02
    • PCT/JP1997003438
    • 1997-09-26
    • LABORATORY OF MOLECULAR BIOPHOTONICSSATO, YoshihiroTSUJI, AkihikoSUGA, Takayuki
    • LABORATORY OF MOLECULAR BIOPHOTONICS
    • C12Q01/68
    • C12Q1/6818
    • Detection probes enabling very convenient and highly accurate and sensitive detection of DNAs or RNAs having specific base sequences in specimens in the presence of the probes in excess of the target nucleic acid; and a detection method. When mixed with a specimen containing the target substance (DNA, RNA, etc.) having a polynucleotide having a specific base sequence, two types of fluorescent label detection probes hybridize, while being adjacent to each other, with the target nucleic acid. As a result, there arises a shift of the resonance energy between the two types of fluorescent dye molecules. When the resonance energy shift occurs in the hybrid thus formed, the fluorescence attenuation of the acceptor fluorescent dye molecule can be sufficiently delayed compared with the fluorescence attenuation of the directly excited acceptor by appropriately regulating (1) the number of bases between the two nucleotides to which the fluorescent dye moleculees have bonded; (2) the structure (double- or single-stranded) of the hybrid between the two nucleotides to which the fluorescent dye molecules have bonded; and (3), on the detection probes, the sites of the nucleotides into which the fluorescent dye molecules are to be introduced; and by (4) using a pair of detection probes with the appropriate selection of the fluorescent dye molecules. The detection method comprises using the above-mentioned detection probes and detecting the target substance by measuring changes in the fluorescence attenuation curve of the acceptor after the irradiation with pulse excitation rays, thus enabling highly accurate and sensitive detection of the target nucleic acid in the presence of the detection probes in excess of the target nucleic acid.
    • 检测探针能够在超过目标核酸的探针存在下,在标本中具有特异性碱基序列的DNA或RNAs的非常方便和高度准确和灵敏的检测; 和检测方法。 当与含有具有特定碱基序列的多核苷酸的靶物质(DNA,RNA等)的样品混合时,两种类型的荧光标记检测探针在彼此相邻的情况下与靶核酸杂交。 结果,在两种类型的荧光染料分子之间产生共振能量的偏移。 当共轭能量偏移发生在如此形成的杂化物中时,与直接激发的受体的荧光衰减相比,受体荧光染料分子的荧光衰减可以通过适当地调节(1)两个核苷酸之间的碱基数 荧光染料分子结合; (2)荧光染料分子键合的两个核苷酸之间的杂交体的结构(双链或单链) 和(3),在检测探针上,将引入荧光染料分子的核苷酸位点; 并且通过(4)使用具有适当选择荧光染料分子的一对检测探针。 检测方法包括使用上述检测探针并通过测量在用脉冲激发射线照射之后受体的荧光衰减曲线的变化来检测目标物质,从而使得能够在存在下高度精确和灵敏地检测靶核酸 的检测探针超过目标核酸。