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    • 1. 发明申请
    • TEST PROBES, COMMON OLIGONUCLEOTIDE CHIPS, NUCLEIC ACID DETECTION METHOD, AND THEIR USES
    • 测试探针,通用寡核苷酸芯片,核酸检测方法及其用途
    • WO2009140802A8
    • 2010-12-09
    • PCT/CN2008001695
    • 2008-10-06
    • SHAAN XI LIFEGEN CO LTDTANG YITONGCHEN CHAOCUI YALIZHU JUANLIYU LONGLINGAO YIWENLI ZHENG
    • TANG YITONGCHEN CHAOCUI YALIZHU JUANLIYU LONGLINGAO YIWENLI ZHENG
    • C40B40/06C12Q1/68C40B20/02C40B20/04
    • C12Q1/6827C12Q2533/107C12Q2525/197
    • High-throughput detection for the interesting base or the mutation site in the nucleic acid sample can be achieved by the linear test probe pairs P1 and P2. The test probe pairs P1 and P2 respectively comprise either of the flanking complementary sequences which are adjacent to the interesting base or the mutation site in the nucleic acid sample. When the test probe pairs P1, P2 are annealed and hybridized to the nucleic acid sample, a gap will be generated at the interesting base or the mutation site position between the probe pairs and the sample. Divide the annealed hybrid sample into four equal reaction systems to which add dATP, dTTP, dCTP, dGTP, respectively. The test probe pairs P1 and P2 will be ligated into one single probe when adding the complementary nucleotide system under the DNA polymerase or ligase. After purified and amplified, the generated single probes are hybridized to the corresponding area in a common oligonucleotide microarray. The generated single probe will give a signal in the hybrid area, and therefore detect and analyze the hybrid signal to determine the base type or the mutation genotype at the detection position. The invention can be applied to the re-sequencing the target nucleic acid sequence, the detection and analysis for the mutation, insertion, or deletion sites of a known nucleic acid sequence, and the genotyping of the pathogenic microorganism.
    • 可以通过线性测试探针对P1和P2来实现核酸样品中感兴趣碱基或突变位点的高通量检测。 测试探针对P1和P2分别包含与感兴趣碱基或核酸样品中突变位点相邻的侧翼互补序列之一。 当测试探针对P1,P2退火并与核酸样品杂交时,将在探针对和样品之间的感兴趣的碱基或突变位点位置产生间隙。 将退火的混合样品分成四个相等的反应体系,分别加入dATP,dTTP,dCTP,dGTP。 当在DNA聚合酶或连接酶下添加互补核苷酸系统时,测试探针对P1和P2将被连接到一个单个探针中。 纯化和扩增后,产生的单个探针与普通寡核苷酸微阵列中的相应区域杂交。 所产生的单个探针将在混合区域中产生信号,因此检测和分析混合信号以确定检测位置的基因型或突变基因型。 本发明可以应用于重新测序目标核酸序列,检测和分析已知核酸序列的突变,插入或缺失位点,以及病原微生物的基因分型。