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    • 25. 发明申请
    • Single Molecule Detection Using Molecular Motors
    • 使用分子电机的单分子检测
    • US20090035751A1
    • 2009-02-05
    • US10582820
    • 2004-12-17
    • Lyian HeWayne Frasch
    • Lyian HeWayne Frasch
    • C12Q1/68
    • C12Q1/6825C12Q1/6834C12Q2565/619C12Q2531/137C12Q2533/107C12Q2565/601C12Q2565/629C12Q2563/125
    • The present invention provides methods and compositions for highly sensitive nucleic acid detection, down to the single nucleic acid molecule level. In one aspect, the present invention provides methods for detecting a target nucleic acid comprising: (a) providing first and second target-specific nucleic acids, wherein the first and second target-specific nucleic acids each comprise sequences complementary to the target nucleic acid; wherein the first target specific nucleic acid is bound to a first affinity tag and the second target-specific nucleic acid is bound to a second affinity tag, wherein the first affinity tag is capable of binding to a molecular motor, and wherein the second affinity tag is capable of binding to a detection probe; (b) contacting the first and second target-specific nucleic acids to a sample under conditions whereby the first and second target-specific nucleic acids will hybridize to the target nucleic acid if the target nucleic acid is present in the sample, wherein upon hybridization to the target nucleic acid the first and second target-specific nucleic acids are directly adjacent to each other; (c) ligating the first and second target-specific nucleic acids together; (d) binding the molecular motor t˜ the first affinity tag and the detection probe to the second affinity tag; (e) inducing movement of the molecular motor; and (f) detecting movement of the molecular motor through the detection probe, wherein the movement of the molecular motor serves to detect the target nucleic acid in the sample.
    • 本发明提供用于高度敏感的核酸检测的方法和组合物,直到单个核酸分子水平。 一方面,本发明提供了用于检测靶核酸的方法,包括:(a)提供第一和第二靶特异性核酸,其中第一和第二靶特异性核酸各自包含与靶核酸互补的序列; 其中所述第一靶特异性核酸与第一亲和标签结合,并且所述第二靶特异性核酸与第二亲和标签结合,其中所述第一亲和标签能够结合分子马达,并且其中所述第二亲和标签 能够结合检测探针; (b)在条件下使第一和第二靶特异性核酸与样品接触,其中如果靶核酸存在于样品中,则第一和第二靶特异性核酸将与靶核酸杂交,其中在与 靶核酸第一和第二靶特异性核酸彼此直接相邻; (c)将第一和第二靶特异性核酸连接在一起; (d)将第一亲和标签和检测探针的分子马达结合到第二亲和标签上; (e)诱导分子马达的运动; 和(f)检测分子马达通过检测探针的运动,其中分子马达的移动用于检测样品中的靶核酸。