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    Composition kits and methods for performing amplification reactions
    1.
    发明授权
    Composition kits and methods for performing amplification reactions 有权
    用于进行扩增反应的组合物试剂盒和方法

    公开(公告)号：US07696337B2

    公开(公告)日：2010-04-13

    申请号：US11574307

    申请日：2005-08-26

    申请人： Michael M. Becker ,  Steven T. Brentano ,  Daniel P. Kolk ,  Wai-Chung Lam ,  Kristin W. Livezey ,  Norman C. Nelson ,  Astrid R. W. Schroder ,  Gary P. Schroth

    发明人： Michael M. Becker ,  Steven T. Brentano ,  Daniel P. Kolk ,  Wai-Chung Lam ,  Kristin W. Livezey ,  Norman C. Nelson ,  Astrid R. W. Schroder ,  Gary P. Schroth

    IPC分类号： C07H21/04 ,  C12Q1/68

    CPC分类号： C12P19/34 ,  C12Q1/6865 ,  C12Q2537/163 ,  C12Q2521/325 ,  C12Q2521/107 ,  C12Q2525/186 ,  C12Q2533/101 ,  C12Q2525/143

    摘要： The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side products. The method uses only one primer, the “priming oligonucleotide,” a 3′blocked promoter oligonucleotide and optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or eliminating the formation of side products. The method of the present invention minimizes or eliminates the emergence of side products, thus providing a high level of specificity. Furthermore, the appearance of side products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or eliminates this problem, thus providing an enhanced level of sensitivity.

    摘要翻译： 本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种 循环在下一个）。 特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少了副产物的外观。 该方法仅使用一种引物，即引物寡核苷酸，3'封闭的启动子寡核苷酸和任选的终止引物延伸反应的手段，以体外扩增RNA或DNA分子，同时减少或消除副产物的形成。 本发明的方法使副产物的出现最小化或消除，从而提供高水平的特异性。 此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。 本发明使这个问题最小化或消除了这一点，从而提供了增强的灵敏度。

    Single-primer nucleic acid amplification methods
    2.
    发明授权
    Single-primer nucleic acid amplification methods 有权
    单引物核酸扩增方法

    公开(公告)号：US07374885B2

    公开(公告)日：2008-05-20

    申请号：US11213519

    申请日：2005-08-26

    申请人： Michael M. Becker ,  Wai-Chung Lam ,  Kristin W. Livezey ,  Steven T. Brentano ,  Daniel P. Kolk ,  Astrid R. W. Schroder

    发明人： Michael M. Becker ,  Wai-Chung Lam ,  Kristin W. Livezey ,  Steven T. Brentano ,  Daniel P. Kolk ,  Astrid R. W. Schroder

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12P19/34 ,  C12Q1/6865 ,  C12Q2537/163 ,  C12Q2521/325 ,  C12Q2521/107 ,  C12Q2525/186 ,  C12Q2533/101 ,  C12Q2525/143

    摘要： The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side-products. The method uses only one primer, the “priming oligonucleotide,” a promoter oligonucleotide modified to prevent polymerase extension from its 3′-terminus and, optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or substantially eliminating the formation of side-products. The method of the present invention minimizes or substantially eliminates the emergence of side-products, thus providing a high level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or substantially eliminates this problem, thus providing an enhanced level of sensitivity.

    摘要翻译： 本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种 循环在下一个）。 特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少副产物的外观。 该方法仅使用一种引物，即引物寡核苷酸，经修饰的启动子寡核苷酸，用于防止聚合酶从其3'末端延伸，以及任选的终止引物延伸反应的手段，以在体外扩增RNA或DNA分子，同时 减少或基本上消除副产物的形成。 本发明的方法使副产物的出现最小化或基本上消除，从而提供高水平的特异性。 此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。 本发明使这个问题最小化或基本消除，从而提供了增强的灵敏度。

    Method for making available a priming oligonucleotide
    3.
    发明授权
    Method for making available a priming oligonucleotide 有权
    提供引物寡核苷酸的方法

    公开(公告)号：US07939260B2

    公开(公告)日：2011-05-10

    申请号：US12389993

    申请日：2009-02-20

    申请人： Michael M. Becker ,  Steven T. Brentano ,  Kristin W. Livezey ,  Norman C. Nelson ,  Gary P. Schroth

    发明人： Michael M. Becker ,  Steven T. Brentano ,  Kristin W. Livezey ,  Norman C. Nelson ,  Gary P. Schroth

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12P19/34 ,  C12Q1/6865 ,  C12Q2537/163 ,  C12Q2521/325 ,  C12Q2521/107 ,  C12Q2525/186 ,  C12Q2533/101 ,  C12Q2525/143

    摘要： The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side-products. The method uses only one primer, the “priming oligonucleotide,” a promoter oligonucleotide modified to prevent polymerase extension from its 3′-terminus and, optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or substantially eliminating the formation of side-products. The method of the present invention minimizes or substantially eliminates the emergence of side-products, thus providing a high level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or substantially eliminates this problem, thus providing an enhanced level of sensitivity.

    摘要翻译： 本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种 循环在下一个）。 特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少副产物的外观。 该方法仅使用一种引物，即引物寡核苷酸，经修饰的启动子寡核苷酸，用于防止聚合酶从其3'末端延伸，以及任选的终止引物延伸反应的手段，以在体外扩增RNA或DNA分子，同时 减少或基本上消除副产物的形成。 本发明的方法使副产物的出现最小化或基本上消除，从而提供高水平的特异性。 此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。 本发明使这个问题最小化或基本消除，从而提供了增强的灵敏度。

    Method for determining the presence of an RNA analyte in a sample using a modified oligonucleotide probe
    7.
    发明授权
    Method for determining the presence of an RNA analyte in a sample using a modified oligonucleotide probe 有权
    使用修饰的寡核苷酸探针测定样品中RNA分析物的存在的方法

    公开(公告)号：US07070925B1

    公开(公告)日：2006-07-04

    申请号：US09565427

    申请日：2000-05-05

    申请人： Michael M. Becker ,  Mehrdad Majlessi ,  Steven T. Brentano

    发明人： Michael M. Becker ,  Mehrdad Majlessi ,  Steven T. Brentano

    IPC分类号： C07H22/04 ,  C12Q1/68

    CPC分类号： C12Q1/6832 ,  C07H21/00 ,  C12Q1/6816 ,  C12Q1/6844 ,  C12Q1/686 ,  C12Q1/6865 ,  C12Q2563/131 ,  C12Q2563/113 ,  C12Q2525/101 ,  C12Q2525/117 ,  C12Q2527/107

    摘要： The present invention concerns oligonucleotides containing one or more modified nucleotides which increase the binding affinity of the oligonucleotides to target nucleic acids having a complementary nucleotide base sequence. These modified oligonucleotides hybridize to the target sequence at a faster rate than unmodified oligonucleotides having an identical nucleotide base sequence. Such modified oligonucleotides include oligonucleotides containing at least one 2′-O-methylribofuranosyl moiety joined to a nitrogenous base. Oligonucleotides can be modified in accordance with the present invention to preferentially bind RNA targets. The present invention also concerns methods of using these modified oligonucleotides and kits containing the same.

    摘要翻译： 本发明涉及含有一个或多个修饰的核苷酸的寡核苷酸，其增加寡核苷酸与具有互补核苷酸碱基序列的靶核酸的结合亲和力。 这些修饰的寡核苷酸以比具有相同核苷酸碱基序列的未修饰寡核苷酸更快的速率与靶序列杂交。 这种修饰的寡核苷酸包括含有至少一个连接到含氮碱基的2'-O-甲基呋喃核糖基部分的寡核苷酸。 可以根据本发明修饰寡核苷酸以优先结合RNA靶。 本发明还涉及使用这些修饰的寡核苷酸的方法和含有该寡核苷酸的试剂盒。

    Kits for amplifying target nucleic acid sequences using modified oligonucleotides
    9.
    发明授权
    Kits for amplifying target nucleic acid sequences using modified oligonucleotides 有权
    使用修饰的寡核苷酸扩增靶核酸序列的试剂盒

    公开(公告)号：US06903206B1

    公开(公告)日：2005-06-07

    申请号：US09523237

    申请日：2000-03-10

    申请人： Michael M. Becker ,  Mehrdad Majlessi ,  Steven T. Brentano

    发明人： Michael M. Becker ,  Mehrdad Majlessi ,  Steven T. Brentano

    IPC分类号： C12N15/09 ,  C07H21/00 ,  C12Q1/68 ,  C07H21/04

    CPC分类号： C07H21/00 ,  C12Q1/6832 ,  C12Q1/686 ,  C12Q1/6865 ,  C12Q2527/107 ,  C12Q2525/113 ,  C12Q2523/113 ,  C12Q2537/101 ,  C12Q2565/107

    摘要： The present invention concerns oligonucleotides containing one or more modified nucleotides which increase the binding affinity of the oligonucleotides to target nucleic acids having a complementary nucleotide base sequence. These modified oligonucleotides hybridize to the target sequence at a faster rate than unmodified oligonucleotides having an identical nucleotide base sequence. Such modified oligonucleotides include oligonucleotides containing at least one 2′-O-methylribofuranosyl moiety joined to a nitrogenous base. Oligonucleotides can be modified in accordance with the present invention to preferentially bind RNA targets. The present invention also concerns methods of using these modified oligonucleotides and kits containing the same.

    摘要翻译： 本发明涉及含有一个或多个修饰的核苷酸的寡核苷酸，其增加寡核苷酸与具有互补核苷酸碱基序列的靶核酸的结合亲和力。 这些修饰的寡核苷酸以比具有相同核苷酸碱基序列的未修饰寡核苷酸更快的速率与靶序列杂交。 这种修饰的寡核苷酸包括含有至少一个连接到含氮碱基的2'-O-甲基呋喃核糖基部分的寡核苷酸。 可以根据本发明修饰寡核苷酸以优先结合RNA靶。 本发明还涉及使用这些修饰的寡核苷酸的方法和含有该寡核苷酸的试剂盒。