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    Methods and compositions for nucleic acid amplification
    1.
    发明授权
    Methods and compositions for nucleic acid amplification 有权
    用于核酸扩增的方法和组合物

    公开(公告)号：US08198027B2

    公开(公告)日：2012-06-12

    申请号：US11962072

    申请日：2007-12-20

    申请人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Norman C. Nelson ,  Lyle J. Arnold, Jr.

    发明人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Norman C. Nelson ,  Lyle J. Arnold, Jr.

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04

    CPC分类号： C12Q1/6881 ,  C12Q1/6834 ,  C12Q1/6844 ,  C12Q1/6865 ,  C12Q2525/155 ,  C12Q2525/186 ,  C12Q2525/197 ,  C12Q2531/143 ,  C12Q2565/543

    摘要： Compositions that are used in nucleic acid amplification in vitro are disclosed, which include a target specific universal (TSU) promoter primer or promoter provider oligonucleotide that includes a target specific (TS) sequence that hybridizes specifically to a target sequence that is amplified and a universal (U) sequence that is introduced into the sequence that is amplified, by using a primer for the universal sequence. Methods of nucleic acid amplification in vitro are disclosed that use one or more TSU oligonucleotides to attached a U sequence to a target nucleic acid in a target capture step and then use a primer for a U sequence in subsequent amplification steps performed in substantially isothermal conditions to make amplification products that contain a U sequence that indicates the presence of the target nucleic acid in a sample.

    摘要翻译： 公开了用于体外核酸扩增的组合物，其包括目标特异性通用（TSU）启动子引物或启动子提供者寡核苷酸，其包括与扩增的靶序列特异性杂交的靶特异性（TS）序列和通用 （U）序列，通过使用通用序列的引物引入扩增的序列。 公开了体外核酸扩增方法，其中使用一种或多种TSU寡核苷酸将U序列连接到目标捕获步骤中的靶核酸，然后在基本上等温条件下进行的随后扩增步骤中使用U序列引物 制备含有指示样品中靶核酸存在的U序列的扩增产物。

    Methods and compositions for nucleic acid amplification
    2.
    发明授权
    Methods and compositions for nucleic acid amplification 有权
    用于核酸扩增的方法和组合物

    公开(公告)号：US09169512B2

    公开(公告)日：2015-10-27

    申请号：US13380018

    申请日：2010-07-01

    申请人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Michael M. Becker ,  Norman C. Nelson ,  Lyle J. Arnold, Jr.

    发明人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Michael M. Becker ,  Norman C. Nelson ,  Lyle J. Arnold, Jr.

    IPC分类号： C12P19/34 ,  C12Q1/68

    CPC分类号： C12Q1/6865 ,  C12Q1/6844 ,  C12Q1/6848 ,  C12Q1/6853 ,  C12Q2549/119 ,  C12Q2537/143 ,  C12Q2531/143

    摘要： Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.

    摘要翻译： 用于进行扩增反应的组合物，反应混合物和方法，包括多重扩增反应，其中所述方法包括使用包含连接的第一和第二扩增寡聚体成员的一种或多种扩增寡聚体复合物。 一方面，将扩增寡聚物复合物与靶核酸杂交，然后捕获具有杂交扩增寡聚物复合物的靶核酸，并洗涤其它组分。 靶核酸的靶序列被预扩增以产生第一扩增产物。 第一扩增产物在一个或多个二级扩增反应中扩增以产生第二扩增产物。

    Methods and compositions for nucleic acid amplification
    3.
    发明授权
    Methods and compositions for nucleic acid amplification 有权
    用于核酸扩增的方法和组合物

    公开(公告)号：US08642268B2

    公开(公告)日：2014-02-04

    申请号：US13460341

    申请日：2012-04-30

    申请人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Norman C. Nelson ,  Lyle J. Arnold, Jr. ,  Michael M. Becker

    发明人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Norman C. Nelson ,  Lyle J. Arnold, Jr. ,  Michael M. Becker

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H19/00

    CPC分类号： C12Q1/6881 ,  C12Q1/6834 ,  C12Q1/6844 ,  C12Q1/6865 ,  C12Q2525/155 ,  C12Q2525/186 ,  C12Q2525/197 ,  C12Q2531/143 ,  C12Q2565/543

    摘要： Compositions that are used in nucleic acid amplification in vitro are disclosed, which include a target specific universal (TSU) promoter primer or promoter provider oligonucleotide that includes a target specific (TS) sequence that hybridizes specifically to a target sequence that is amplified and a universal (U) sequence that is introduced into the sequence that is amplified, by using a primer for the universal sequence. Methods of nucleic acid amplification in vitro are disclosed that use one or more TSU oligonucleotides to attached a U sequence to a target nucleic acid in a target capture step and then use a primer for a U sequence in subsequent amplification steps performed in substantially isothermal conditions to make amplification products that contain a U sequence that indicates the presence of the target nucleic acid in a sample.

    摘要翻译： 公开了用于体外核酸扩增的组合物，其包括目标特异性通用（TSU）启动子引物或启动子提供者寡核苷酸，其包括与扩增的靶序列特异性杂交的靶特异性（TS）序列和通用 （U）序列，通过使用通用序列的引物引入扩增的序列。 公开了体外核酸扩增方法，其中使用一种或多种TSU寡核苷酸将U序列连接到目标捕获步骤中的靶核酸，然后在基本上等温条件下进行的随后扩增步骤中使用U序列引物 制备含有指示样品中靶核酸存在的U序列的扩增产物。

    METHODS AND COMPOSITIONS FOR NUCLEIC ACID AMPLIFICATION
    5.
    发明申请
    METHODS AND COMPOSITIONS FOR NUCLEIC ACID AMPLIFICATION 有权
    用于核酸扩增的方法和组合物

    公开(公告)号：US20120178636A1

    公开(公告)日：2012-07-12

    申请号：US13380018

    申请日：2010-07-01

    申请人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Michael M. Becker ,  Norman C. Nelson ,  Lyle J. Arnold, JR.

    发明人： Steven T. Brentano ,  Dmitry Lyakhov ,  James D. Carlson ,  Michael M. Becker ,  Norman C. Nelson ,  Lyle J. Arnold, JR.

    IPC分类号： C40B30/00

    CPC分类号： C12Q1/6865 ,  C12Q1/6844 ,  C12Q1/6848 ,  C12Q1/6853 ,  C12Q2549/119 ,  C12Q2537/143 ,  C12Q2531/143

    摘要： Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.

    摘要翻译： 用于进行扩增反应的组合物，反应混合物和方法，包括多重扩增反应，其中所述方法包括使用包含连接的第一和第二扩增寡聚体成员的一种或多种扩增寡聚体复合物。 一方面，将扩增寡聚物复合物与靶核酸杂交，然后捕获具有杂交扩增寡聚物复合物的靶核酸，并洗涤其它组分。 靶核酸的靶序列被预扩增以产生第一扩增产物。 第一扩增产物在一个或多个二级扩增反应中扩增以产生第二扩增产物。

    Homogeneous protection assay
    8.
    发明授权
    Homogeneous protection assay 失效
    均质保护试验

    公开(公告)号：US5639604A

    公开(公告)日：1997-06-17

    申请号：US161706

    申请日：1993-12-03

    申请人： Lyle J. Arnold, Jr. ,  Norman C. Nelson

    发明人： Lyle J. Arnold, Jr. ,  Norman C. Nelson

    IPC分类号： C12Q1/68 ,  G01N33/53 ,  G01N33/532 ,  G01N33/533 ,  G01N33/542 ,  G01N33/58 ,  C07H21/04 ,  C12Q1/00

    CPC分类号： G01N33/58 ,  C12Q1/6816 ,  C12Q1/6823 ,  G01N33/5306 ,  G01N33/532 ,  G01N33/533 ,  G01N33/542

    摘要： Improved homogenous diagnostic assay methods and labels for detecting an analyte in a medium when the analyte is a member of a specific binding pair. The methods and labels provide procedures for reducing background and increasing sensitivity. The binding partner of the analyte is labeled with a substance, the stability of which detectably changes whenever said analyte is bound as a member of the specific binding pair. In a closely related system, the analyte is labeled with a substance susceptible to differential degradation depending on whether or not the analyte is bound as a member of its specific binding pair. After incubation and selective degradation or chemical or biochemical alteration, the amount of analyte bound is detected by measuring either the stability change or the extent of degradation of the label. In a particular system, chemiluminescent acridinium ester labeled probes are used in a homogenous hybridization assay format for sensitively detecting the presence of complement any target polynucleotide sequences.

    摘要翻译： 当分析物是特异性结合对的成员时，改进的同源诊断测定方法和用于检测培养基中分析物的标记。 方法和标签提供了降低背景和提高灵敏度的程序。 分析物的结合配偶体用物质标记，当所述分析物作为特异性结合对的成员结合时，其稳定性可检测地改变。 在紧密相关的系统中，根据分析物是否作为其特异性结合对的成员结合，分析物被易受差异降解敏感的物质标记。 孵育和选择性降解或化学或生物化学改变后，通过测量标签的稳定性变化或降解程度来检测分析物的结合量。 在特定系统中，化学发光的吖啶酯标记的探针以均质杂交测定形式使用，以敏感地检测补体存在任何靶多核苷酸序列。

    Acridinium ester labelling and purification of nucleotide probes
    9.
    发明授权
    Acridinium ester labelling and purification of nucleotide probes 失效
    吖啶酯标记和纯化核苷酸探针

    公开(公告)号：US5185439A

    公开(公告)日：1993-02-09

    申请号：US332939

    申请日：1988-12-12

    申请人： Lyle J. Arnold, Jr. ,  Norman C. Nelson

    发明人： Lyle J. Arnold, Jr. ,  Norman C. Nelson

    IPC分类号： C07H21/00

    CPC分类号： C07H21/00

    摘要： Methods for the construction, labelling and subsequent purification of nucleic acid probes containing primary amines with acridinium esters 4-(2-succinimidyloxycarbonyl-ethyl)phenyl-10-methylacridinium-9-carboxylate fluorosulfonate). The method for attaching acridinium esters to probes uses high (0.1 to 50 mM) acridinium ester concentrations achieved using organic solvent in concentrations of 20% to 80% by volume, and may be carried out either in solution, or with one or the other of the acridinium ester or the probe suspended in solution. Purification (the separation of labelled probe from unlabelled probe and free label) involves (1) first removing most of the free acridinium ester label from probe using rapid separation techniques, then (2) removing substantially all remaining free label from the probe and separating labelled probe from unlabelled probe, involves specific applications of ion exchange, reverse phase or hydroxyapatite HPLC.

    摘要翻译： 含有伯胺与吖啶酯4-（2-琥珀酰亚氨基氧羰基 - 乙基）苯基-10-甲基吖啶-9-甲酸氟代磺酸酯的核酸探针的构建，标记和随后纯化的方法。 将吖啶酯连接到探针的方法使用浓度为20％至80％（体积）的有机溶剂获得的高（0.1至50mM）吖啶酯浓度，并且可以在溶液中或与其中的一种或一种 吖啶酯或探针悬浮在溶液中。 纯化（标记的探针与未标记的探针和游离标记物的分离）包括：（1）首先使用快速分离技术从探针中除去大部分游离的吖啶酯标记物，然后（2）从探针中基本上除去所有剩余的游离标记并分离标记 来自未标记探针的探针涉及离子交换，反相或羟基磷灰石HPLC的具体应用。