公开(公告)号：US07667024B2

公开(公告)日：2010-02-23

申请号：US10993625

申请日：2004-11-19

申请人： Fei Mao ,  Xing Xin

发明人： Fei Mao ,  Xing Xin

IPC分类号： C07H21/02 ,  C07H21/04 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6818 ,  C12Q1/6851 ,  Y10S435/975 ,  C12Q2565/1015 ,  C12Q2561/101 ,  C12Q2565/1025

摘要： An embodiment of the invention discloses new methods for designing labeled nucleic acid probes and primers by labeling oligonucleotides with a plurality of spectrally identical or similar dyes and optionally with one or more quencher dyes. Oligonucleotides labeled in accordance with some embodiments of the invention exhibit a detectable increase in signal, for example, fluorescent signal when the labeling dyes are separated from one another. Methods for separating the dye include cleaving the labeled oligonucleotides include using enzymes that have 5′-exonuclease activity. In one embodiment nucleic acid primers of the present invention may fluoresce upon hybridization to a target sequence and incorporation into the amplification product. Nucleic acid probes and primers of the present invention have wide applications ranging from general detection of a target nucleic acid sequence to clinical diagnostics. Major advantages of the oligonucleotides including nucleic acid probes and primers of many embodiments of the present invention are their synthetic simplicity, spectral versatility and superior fluorescent signal.

摘要翻译： 本发明的一个实施方案公开了通过用多个光谱相同或相似的染料和任选地与一种或多种猝灭剂染料标记寡核苷酸来设计标记的核酸探针和引物的新方法。 根据本发明的一些实施方案标记的寡核苷酸在标记染料彼此分离时表现出可检测的信号增加，例如荧光信号。 用于分离染料的方法包括切割标记的寡核苷酸包括使用具有5'-外切核酸酶活性的酶。 在一个实施方案中，本发明的核酸引物可以在与靶序列杂交时发荧光，并掺入扩增产物。 本发明的核酸探针和引物具有广泛的应用范围，从靶核酸序列的一般检测到临床诊断。 包括本发明许多实施方案的核酸探针和引物的寡核苷酸的主要优点是它们的合成简便性，光谱通用性和优异的荧光信号。