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    Method of detecting specific substances by selective growth of living cells
    1.
    发明授权
    Method of detecting specific substances by selective growth of living cells 失效
    通过活细胞选择性生长检测特定物质的方法

    公开(公告)号：US5085982A

    公开(公告)日：1992-02-04

    申请号：US873504

    申请日：1986-06-12

    申请人： Douglas H. Keith

    发明人： Douglas H. Keith

    IPC分类号： C12Q1/02 ,  C12Q1/68 ,  C12Q1/70

    CPC分类号： C12Q1/70 ,  C12Q1/02 ,  C12Q1/682

    摘要： The invented method utilizes a signal amplification system comprising living cells which are specifically provided with the ability to survive, reproduce and be detected in the event that a target molecule is present. The method comprises first the step of binding a target molecule to a substratum. In the second step a phagemid is prepared which is capable of transfecting a cell enabling such transfected cell to produce a signal, such as color or light. The phagemid is then provided with a means for binding to a probe, thus forming a phagemid complex.The probe may also be provided with a means of binding to the phagemid complex. This modified probe is then hybridized with the target molecules bound to the substratum. Thereafter, the phagemid complex is hybridized with the modified probe and specific binding occurs between the phagemid complex and the modified probe thus forming a substrutum-target-probe phagemid complex. As a next step, lysogenic bacterial cells which are infectable by the particular phagemid being used are added to the system in an environment which favors transfection of the phagemid nucleic acid into the bacteria. The growth medium is constituted so that only transfected bacteria survive and are capable of reproducing. The phagemid also enable the bacteria to produce a signal such light or color. Only the transfected bacterial cells containing the plasmid gene for producing the signal will be detectable.

    摘要翻译： 本发明的方法利用了包含活细胞的信号放大系统，该活细胞在靶分子存在的情况下特异性地具有存活，繁殖和检测的能力。 该方法首先包括将靶分子与底物结合的步骤。 在第二步中，制备能够转染细胞的噬菌粒，使得这样的转染细胞能够产生诸如颜色或光的信号。 然后向噬菌粒提供与探针结合的方法，从而形成噬菌粒复合物。 探针也可以具有结合噬菌体复合物的方法。 然后将该修饰的探针与结合到底物的靶分子杂交。 此后，噬菌粒复合物与修饰的探针杂交，并且在噬菌粒复合物和修饰的探针之间发生特异性结合，从而形成子目标 - 探针噬菌粒复合物。 作为下一步骤，在有利于将噬菌粒核酸转染到细菌的环境中，将可被特定噬菌粒感染的溶原性细菌细胞加入到该系统中。 构建生长培养基使得只有转染的细菌才能存活并能够繁殖。 噬菌粒还能使细菌产生如光或颜色的信号。 只有含有用于产生该信号的质粒基因的转染细菌细胞将是可检测的。

    Nucleic acid detection with separation
    3.
    发明授权
    Nucleic acid detection with separation 失效
    分离核酸检测

    公开(公告)号：US6054266A

    公开(公告)日：2000-04-25

    申请号：US276139

    申请日：1988-11-23

    申请人： Mel N. Kronick ,  Douglas H. Keith ,  Lincoln J. McBride ,  Norman M. Whiteley ,  Michael W. Hunkapiller

    发明人： Mel N. Kronick ,  Douglas H. Keith ,  Lincoln J. McBride ,  Norman M. Whiteley ,  Michael W. Hunkapiller

    IPC分类号： G01N33/50 ,  C12Q1/68 ,  G01N30/88 ,  G01N33/58 ,  C07H21/04

    CPC分类号： C12Q1/6813 ,  C12Q1/6804 ,  C12Q1/6823

    摘要： Nucleic acid sequences are detected by a multi-step process, involving labeling sample nucleic acid sequences, duplexing the labeled sample with a probe having a coupling element, immobilizing all of the duplexed probe and target sequence and unduplexed probe, separating specifically immobilized nucleic acid from free and non-specifically immobilized nucleic acid, releasing specifically immobilized nucleic acid, and detecting the presence of the sequence of interest by means of the label. The labeled sequence may be characterized by sizing, e.g. electrophoresis. The method provides for a sensitive and rapid means for accurate detection of sequences of interest in a wide variety of situations.

    摘要翻译： 通过多步法检测核酸序列，包括标记样品核酸序列，用具有偶联元件的探针双链化标记的样品，固定所有双链体探针和靶序列和未折射的探针，将特异性固定的核酸与 游离和非特异性固定化的核酸，释放特异性固定化的核酸，并通过标记物检测感兴趣的序列的存在。 标记序列的特征可以是尺寸，例如 电泳。 该方法提供用于在各种情况下精确检测感兴趣序列的敏感且快速的方法。