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公开(公告)号：US20110014617A1
公开(公告)日：2011-01-20
申请号：US12816006
申请日：2010-06-15
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6834 , C12Q1/6823 , C12Q1/6825 , C12Q2531/113 , C12Q2521/307 , C12Q2525/204 , C12Q2565/607 , C12Q2561/109 , C12Q2565/519
摘要： Methods and kits for detecting a target nucleic acid in a sample are described. In some embodiments, the sample to be analyzed includes a primer which hybridizes to at least a portion of the target nucleic acid, a probe having a first region which hybridizes to at least a portion of the target nucleic acid and a second region having a detectable label, a polymerase which extends the hybridized primer and an enzyme comprising nuclease activity that can cleave the hybridized hybridization probe to thereby release a labeled probe fragment. In some embodiments, the sample can then be contacted with a solid support comprising surface bound capture probes which can hybridize to the labeled probe fragment(s). These capture probes more readily bind to the probe fragment(s) than to the intact hybridization probe. The label can then be detected on the support surface. In this manner, improved discrimination between the probe fragments and the intact hybridization probes can be achieved.
摘要翻译： 描述了用于检测样品中靶核酸的方法和试剂盒。 在一些实施方案中，待分析的样品包括与靶核酸的至少一部分杂交的引物，具有与靶核酸的至少一部分杂交的第一区的探针和具有可检测的第二区 标记，扩增杂交引物的聚合酶和包含核酸酶活性的酶，其可以切割杂交的杂交探针，从而释放标记的探针片段。 在一些实施方案中，然后可以将样品与包含可以与标记的探针片段杂交的表面结合捕获探针的固体支持物接触。 这些捕获探针比完整的杂交探针更容易与探针片段结合。 然后可以在支撑表面上检测标签。 以这种方式，可以实现探针片段与完整杂交探针之间的改善的区别。
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公开(公告)号：US20080193940A1
公开(公告)日：2008-08-14
申请号：US11965807
申请日：2007-12-28
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6823 , C12Q1/6825 , C12Q1/6827 , C12Q2531/113 , C12Q2521/307 , C12Q2525/161 , C12Q2525/301 , C12Q2565/607 , C12Q2565/519 , C12Q2521/301 , C12Q2561/109
摘要： A method and kit for detecting a target nucleic acid in a sample is described. The sample to be analyzed may include a primer which hybridizes to at least a portion of the target nucleic acid, a probe having a first region which hybridizes to at least a portion of the target nucleic acid and a second region having a detectable label, a polymerase which extends the hybridized primer and an enzyme comprising exonuclease activity that can cleave the hybridized hybridization probe to thereby generate a labeled probe fragment. At least one portion of the hybridization probe hybridizes to another portion of the hybridization probe to thereby form a folded structure. The method can involve melting the sample, reducing the temperature of the sample to allow primer and probe to each hybridize to at least a portion of single stranded target nucleic acid in the sample, elongating the primer and releasing the labeled probe fragment. The sample can be contacted with a solid support comprising surface bound capture probes which hybridize to the labeled probe fragments. The label can then be detected.
摘要翻译： 描述了用于检测样品中靶核酸的方法和试剂盒。 要分析的样品可以包括与靶核酸的至少一部分杂交的引物，具有与靶核酸的至少一部分杂交的第一区的探针和具有可检测标记的第二区， 扩增杂交引物的聚合酶和包含外切核酸酶活性的酶，其可以切割杂交的杂交探针，从而产生标记的探针片段。 至少一部分杂交探针与杂交探针的另一部分杂交，从而形成折叠结构。 该方法可以包括熔化样品，降低样品的温度以使引物和探针各自与样品中的至少一部分单链靶核酸杂交，延长引物并释放标记的探针片段。 样品可以与包含与标记的探针片段杂交的表面结合捕获探针的固体支持物接触。 然后可以检测标签。
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公开(公告)号：US20080241838A1
公开(公告)日：2008-10-02
申请号：US11965837
申请日：2007-12-28
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6834 , C12Q1/6823 , C12Q1/6825 , C12Q2531/113 , C12Q2521/307 , C12Q2525/204 , C12Q2565/607 , C12Q2561/109 , C12Q2565/519
摘要： Methods and kits for detecting a target nucleic acid in a sample are described. In some embodiments, the sample to be analyzed includes a primer which hybridizes to at least a portion of the target nucleic acid, a probe having a first region which hybridizes to at least a portion of the target nucleic acid and a second region having a detectable label, a polymerase which extends the hybridized primer and an enzyme comprising nuclease activity that can cleave the hybridized hybridization probe to thereby release a labeled probe fragment. In some embodiments, the sample can then be contacted with a solid support comprising surface bound capture probes which can hybridize to the labeled probe fragment(s). These capture probes more readily bind to the probe fragment(s) than to the intact hybridization probe. The label can then be detected on the support surface. In this manner, improved discrimination between the probe fragments and the intact hybridization probes can be achieved.
摘要翻译： 描述了用于检测样品中靶核酸的方法和试剂盒。 在一些实施方案中，待分析的样品包括与靶核酸的至少一部分杂交的引物，具有与靶核酸的至少一部分杂交的第一区的探针和具有可检测的第二区 标记，扩增杂交引物的聚合酶和包含核酸酶活性的酶，其可以切割杂交的杂交探针，从而释放标记的探针片段。 在一些实施方案中，然后可以将样品与包含可以与标记的探针片段杂交的表面结合捕获探针的固体支持物接触。 这些捕获探针比完整的杂交探针更容易与探针片段结合。 然后可以在支撑表面上检测标签。 以这种方式，可以实现探针片段与完整杂交探针之间的改善的区别。
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4.发明申请Compositions, methods, and kits for selective amplification of nucleic acids 审中-公开用于选择性扩增核酸的组合物，方法和试剂盒公开(公告)号：US20070212695A1
公开(公告)日：2007-09-13
申请号：US11331599
申请日：2006-01-12
CPC分类号： C12Q1/6827 , C12Q1/6844 , C12Q1/6853 , Y02A50/54 , C12Q2525/307 , C12Q2531/125 , C12Q2525/117 , C12Q2561/125 , C12Q2537/143
摘要： The current teachings are directed to compositions, methods, and kits for selectively amplifying and for detecting target sequences. In some embodiments, a circularizable probe and/or a probe pair are disclosed for selectively amplifying target sequences. Methods for selectively amplifying target sequences are also disclosed, as are methods for detecting selectively amplified target sequences. Certain embodiments of the disclosed methods comprise a circularizable probe, a probe pair, comprising a first probe and a second probe, or both. In certain embodiments, a multiplicity of different circularizable probes, a multiplicity of different probe sets, or a multiplicity of different circularizable probes and a multiplicity of different probe sets are provided to selectively amplify or to detect a multiplicity of different target sequences, typically in a multiplex reaction. According to certain disclosed methods, surrogates of the target sequences are selectively amplified, including without limitation ligated probes, first amplification products, second amplification products, or combinations thereof. In some embodiments, selectively amplified target sequences or their surrogates are detected, directly or indirectly, indicating the presence of the corresponding target sequence. Kits to facilitate the performance of the disclosed methods are also provided.
摘要翻译： 目前的教导涉及用于选择性扩增和检测靶序列的组合物，方法和试剂盒。 在一些实施方案中，公开了可圆化探针和/或探针对，用于选择性扩增靶序列。 还公开了选择性扩增靶序列的方法，以及用于检测选择性扩增的靶序列的方法。 所公开方法的某些实施方案包括可圆化的探针，包括第一探针和第二探针的探针对，或两者。 在某些实施方案中，提供多个不同的可圆化探针，多个不同的探针组或多个不同的可环化探针和多个不同的探针组，以选择性地扩增或检测多种不同靶序列，通常在 多重反应。 根据某些公开的方法，选择性扩增靶序列的替代物，包括但不限于连接的探针，第一扩增产物，第二扩增产物或其组合。 在一些实施方案中，直接或间接检测选择性扩增的靶序列或其替代物，表明存在相应的靶序列。 还提供了用于促进所公开方法的性能的套件。
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5.发明申请Compositions, Methods, and Kits for Selective Amplification of Nucleic Acids 审中-公开用于核酸选择性扩增的组合物，方法和试剂盒公开(公告)号：US20100184159A1
公开(公告)日：2010-07-22
申请号：US12692557
申请日：2010-01-22
IPC分类号： C12P19/34
CPC分类号： C12Q1/6827 , C12Q1/6844 , C12Q1/6853 , Y02A50/54 , C12Q2525/307 , C12Q2531/125 , C12Q2525/117 , C12Q2561/125 , C12Q2537/143
摘要： The current teachings are directed to compositions, methods, and kits for selectively amplifying and for detecting target sequences. In some embodiments, a circularizable probe and/or a probe pair are disclosed for selectively amplifying target sequences. Methods for selectively amplifying target sequences are also disclosed, as are methods for detecting selectively amplified target sequences. Certain embodiments of the disclosed methods comprise a circularizable probe, a probe pair, comprising a first probe and a second probe, or both. In certain embodiments, a multiplicity of different circularizable probes, a multiplicity of different probe sets, or a multiplicity of different circularizable probes and a multiplicity of different probe sets are provided to selectively amplify or to detect a multiplicity of different target sequences, typically in a multiplex reaction. According to certain disclosed methods, surrogates of the target sequences are selectively amplified, including without limitation ligated probes, first amplification products, second amplification products, or combinations thereof. In some embodiments, selectively amplified target sequences or their surrogates are detected, directly or indirectly, indicating the presence of the corresponding target sequence. Kits to facilitate the performance of the disclosed methods are also provided.
摘要翻译： 目前的教导涉及用于选择性扩增和检测靶序列的组合物，方法和试剂盒。 在一些实施方案中，公开了可圆化探针和/或探针对，用于选择性扩增靶序列。 还公开了选择性扩增靶序列的方法，以及用于检测选择性扩增的靶序列的方法。 所公开方法的某些实施方案包括可圆化的探针，包括第一探针和第二探针的探针对，或两者。 在某些实施方案中，提供多个不同的可圆化探针，多个不同的探针组或多个不同的可环化探针和多个不同的探针组，以选择性地扩增或检测多种不同靶序列，通常在 多重反应。 根据某些公开的方法，选择性扩增靶序列的替代物，包括但不限于连接的探针，第一扩增产物，第二扩增产物或其组合。 在一些实施方案中，直接或间接检测选择性扩增的靶序列或其替代物，表明存在相应的靶序列。 还提供了用于促进所公开方法的性能的套件。
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公开(公告)号：US20070099211A1
公开(公告)日：2007-05-03
申请号：US11488439
申请日：2006-07-17
申请人： Vissarion Aivazachvili , Kristian Scaboo , Aldrich Lau , Konrad Faulstich , Robert Eason , John Van Camp , Timothy Liu
发明人： Vissarion Aivazachvili , Kristian Scaboo , Aldrich Lau , Konrad Faulstich , Robert Eason , John Van Camp , Timothy Liu
CPC分类号： C12Q1/6823 , B01L3/5027 , B01L7/525 , B01L2200/10 , B01L2300/0645 , B01L2300/0681 , B01L2300/1844 , B01L2300/185 , B01L2300/1872 , B01L2300/1894 , B01L2400/0418 , C12Q1/6825 , C12Q2565/607 , C12Q2563/131 , C12Q2565/629 , C12Q2531/113 , C12Q2521/301 , C12Q2523/107
摘要： Methods for detecting a target polynucleotide sequences are provided that utilize a probe having a target-complementary segment and a detectable tag. By cleaving the detectable tab and associating the tag with a tag complement coupled to an electrode, an electrochemical signal can be detected that is related to the presence of the tag:tag complement complex.
摘要翻译： 提供了检测靶多核苷酸序列的方法，其利用具有靶互补区段和可检测标签的探针。 通过切割可检测的标签并将标签与耦合到电极的标签补体相关联，可以检测到与标签：标签补体复合物的存在相关的电化学信号。
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