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    Method and kit involving displacement and rehybridization of labeled polynucleotide
    1.
    发明授权
    Method and kit involving displacement and rehybridization of labeled polynucleotide 失效
    标记多核苷酸置换和再杂交的方法和试剂盒

    公开(公告)号：US4818680A

    公开(公告)日：1989-04-04

    申请号：US811034

    申请日：1985-12-18

    申请人： Mary Collins ,  Joseph P. Dougherty ,  Marian S. Ellwood ,  Edward F. Fritsch ,  Kenneth A. Jacobs

    发明人： Mary Collins ,  Joseph P. Dougherty ,  Marian S. Ellwood ,  Edward F. Fritsch ,  Kenneth A. Jacobs

    IPC分类号： C12Q1/68 ,  G01N33/566

    CPC分类号： C12Q1/6876 ,  C12Q1/6813 ,  C12Q1/6823 ,  Y10S435/81

    摘要： A diagnostic reagent is disclosed which is capable of binding to a target nucleotide sequence which is bound to a labeled polynucleotide in a target binding region which is at least partially co-extensive with the target binding region in the probe polynucleotide which is capable of binding to the target nucleotide sequence. A method is disclosed in which the reagent is contacted with a sample and with a capturing polynucleotide under conditions such that target nucleotide which may be present in the sample binds to the probe polynucleotide and displaces labeled polynucleotide from the reagent complex, and the capturing polynucleotide binds selectively to the displaced labeled polynucleotide in the region of the labeled polynucleotide that had been bound to the probe polynucleotide. Determination of the displaced nucleotide gives a value which is a function of the presence and concentration of target nucleotide in the sample.

    摘要翻译： 公开了一种诊断试剂，其能够结合靶向结合区域中的标记的多核苷酸的靶核苷酸序列，所述靶结合区域与探针多核苷酸中的靶结合区域至少部分共同扩增，所述靶结合区域能够结合 靶核苷酸序列。 公开了一种方法，其中试剂与样品和捕获多核苷酸在条件下接触，使得可能存在于样品中的靶核苷酸与探针多核苷酸结合并从试剂复合物置换标记的多核苷酸，并且捕获多核苷酸结合 选择性地与已经与探针多核苷酸结合的标记的多核苷酸的区域中的置换的标记的多核苷酸。 取代的核苷酸的测定给出一个值，该值是样品中靶核苷酸的存在和浓度的函数。

    Displacement polynucleotide assay method and polynucleotide complex reagent therefor
    2.
    发明授权
    Displacement polynucleotide assay method and polynucleotide complex reagent therefor 失效
    位移多核苷酸测定方法和多核苷酸复合试剂

    公开(公告)号：US4766062A

    公开(公告)日：1988-08-23

    申请号：US607885

    申请日：1984-05-07

    申请人： Steven E. Diamond ,  Joseph G. Brewen ,  Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch

    发明人： Steven E. Diamond ,  Joseph G. Brewen ,  Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch

    IPC分类号： G01N33/50 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/58 ,  G01N33/566

    CPC分类号： C12Q1/6823 ,  C12Q1/6813 ,  C12Q1/6827 ,  C12Q1/6832 ,  C12Q1/6876 ,  Y10S435/803 ,  Y10S435/81

    摘要： A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) contains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the target nucleotide sequence (G) causes binding, initially between G and a single-stranded portion (IBR) of the target binding region (TBR). Thereafter the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample.

    摘要翻译： 公开了一种诊断试剂，其含有通过嘌呤/嘧啶氢键与标记的多核苷酸（L）结合的探针多核苷酸（P）的复合物。 探针（P）含有能够结合生物样品的靶核苷酸序列（G）的靶结合区（TBR）。 公开了一种与含有靶核苷酸序列（G）的样品的接触引起最初在G与靶结合区（TBR）的单链部分（IBR）之间的结合。 此后，通过（G）的分支迁移进入（P）/（L）结合区域，标记的多核苷酸（L）从络合物中移位。 置换的标记多核苷酸（L）的测定给出了样品中靶核苷酸序列（G）的存在和浓度的函数值。

    Displacement polynucleotide assay employing polyether and diagnostic kit
    3.
    发明授权
    Displacement polynucleotide assay employing polyether and diagnostic kit 失效
    使用聚醚和诊断试剂盒的位移多核苷酸测定

    公开(公告)号：US4766064A

    公开(公告)日：1988-08-23

    申请号：US684308

    申请日：1984-12-20

    申请人： Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch ,  Joseph G. Brewen ,  Steven E. Diamond

    发明人： Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch ,  Joseph G. Brewen ,  Steven E. Diamond

    IPC分类号： C12Q1/68 ,  G01N33/566

    CPC分类号： C12Q1/6832 ,  C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6876 ,  Y10S435/803 ,  Y10S435/81

    摘要： A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) contains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the target nucleotide sequence (G) causes binding, initially between G and a single-stranded portion (IBR) of the target binding region (TBR). Thereafter the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. A volume excluding polymeric agent such as poly(ethylene oxide) (PEO or PEG) or other polyethers enhances the rate of appearance of displaced labeled polynucleotide. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample.

    摘要翻译： 公开了一种诊断试剂，其含有通过嘌呤/嘧啶氢键与标记的多核苷酸（L）结合的探针多核苷酸（P）的复合物。 探针（P）含有能够结合生物样品的靶核苷酸序列（G）的靶结合区（TBR）。 公开了一种与含有靶核苷酸序列（G）的样品的接触引起最初在G与靶结合区（TBR）的单链部分（IBR）之间的结合。 此后，通过（G）的分支迁移进入（P）/（L）结合区域，标记的多核苷酸（L）从络合物中移位。 不包括聚合物如聚（环氧乙烷）（PEO或PEG）或其他聚醚的体积增加了置换的标记多核苷酸的出现速率。 置换的标记多核苷酸（L）的测定给出了样品中靶核苷酸序列（G）的存在和浓度的函数值。

    Process and nucleic acid construct for producing reagent complexes useful in determining target nucleotide sequences
    4.
    发明授权
    Process and nucleic acid construct for producing reagent complexes useful in determining target nucleotide sequences 失效
    用于产生可用于测定靶核苷酸序列的试剂复合物的方法和核酸构建体

    公开(公告)号：US5268266A

    公开(公告)日：1993-12-07

    申请号：US938201

    申请日：1986-06-09

    申请人： Edward F. Fritsch ,  Mary Collins

    发明人： Edward F. Fritsch ,  Mary Collins

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

    CPC分类号： C12Q1/6827 ,  C12Q1/6813 ,  C12Q1/6823 ,  C12Q1/683 ,  C12Q1/6832 ,  C12Q1/6876

    摘要： A nucleic acid construct useful in preparing reagents for determining target nucleotide sequences in the nucleic acid of a biological sample, the construct having in its single-stranded form:(a) a target binding region substantially complementary to the target nucleotide sequence, and(b) a signal strand pairing segment bound in the construct by complementary base pairing to a portion of the target binding region;a second portion of the target binding region being single-stranded; andthe target binding region and signal strand pairing segment being covalently linked by a phosphate/sugar backbone.A replicable nucleic acid having an origin of replication and two half-restriction sites capable of forming a restriction site can be treated with a restriction enzyme to form a length of nucleic acid containing the target binding region and the signal strand pairing segment. Subsequent labeling of the construct and various optional cleavage and derivation steps can convert the construct to a reagent complex.

    摘要翻译： PCT No.PCT / US86 / 00742 Sec。 371日期1986年6月9日第 102（e）日期1986年6月9日PCT提交1986年4月11日PCT公布。 公开号WO86 / 06412 日期：1986年11月6日。一种用于制备用于测定生物样品核酸中的靶核苷酸序列的试剂的核酸构建体，所述构建体具有单链形式：（a）与 靶核苷酸序列，和（b）通过与靶结合区域的一部分的互补碱基配对结合在构建体中的信号链配对区段; 目标结合区域的第二部分是单链的; 并且靶结合区和信号链配对片段由磷酸/糖主链共价连接。 可以用限制酶处理具有复制起点的复制核酸和两个能够形成限制性位点的半限制性位点，以形成一定长度的含有靶结合区和信号链配对片段的核酸。 随后标记构建体和各种任选的切割和衍生步骤可将构建体转化为试剂复合物。

    Reagent polynucleotide complex with multiple target binding regions, and kit and methods
    5.
    发明授权
    Reagent polynucleotide complex with multiple target binding regions, and kit and methods 失效
    具有多个靶结合区的试剂多核苷酸复合物，试剂盒和方法

    公开(公告)号：US4725536A

    公开(公告)日：1988-02-16

    申请号：US777796

    申请日：1985-09-19

    申请人： Edward F. Fritsch ,  Mary Collins

    发明人： Edward F. Fritsch ,  Mary Collins

    IPC分类号： C12Q1/68 ,  G01N33/566

    CPC分类号： C12Q1/6883 ,  C12Q1/6823 ,  C12Q1/6827 ,  Y10S435/81

    摘要： Reagent complexes containing a probe polynucleotide with at least two target binding regions. A labeled polynucleotide is bound to at least a portion of each target binding region. In one method, sample nucleic acid displaces labeled polynucleotide from one target binding region; after washing, a reagent polynucleotide displaces labeled polynucleotide from the other target binding region. In a second method, sample nucleic acid strands having two target nucleic acid sequences in proximity (e.g., sequences translocated in certain cancers) displace the labeled polynucleotide from both target binding region.

    摘要翻译： 含有具有至少两个靶结合区的探针多核苷酸的试剂复合物。 标记的多核苷酸与每个靶结合区域的至少一部分结合。 在一种方法中，样品核酸从一个靶结合区域置换标记的多核苷酸; 洗涤后，试剂多核苷酸将标记的多核苷酸从其他靶结合区置换。 在第二种方法中，具有接近的两个靶核酸序列的样品核酸链（例如在某些癌症中易位的序列）从两个靶结合区置换标记的多核苷酸。

    Displacement polynucleotide method and reagent complex employing labeled probe polynucleotide
    6.
    发明授权
    Displacement polynucleotide method and reagent complex employing labeled probe polynucleotide 失效
    位移多核苷酸方法和使用标记的探针多核苷酸的试剂复合物

    公开(公告)号：US4752566A

    公开(公告)日：1988-06-21

    申请号：US809992

    申请日：1985-12-17

    申请人： Mary Collins ,  Joseph P. Dougherty

    发明人： Mary Collins ,  Joseph P. Dougherty

    IPC分类号： C12Q1/68 ,  G01N33/566

    CPC分类号： C12Q1/6823 ,  C12Q1/6816 ,  C12Q1/6834

    摘要： A method for determining the presence and amount of a predetermined target nucleotide sequence in a biological sample is provided. The method employs a reagent complex which consists of a labeled probe polynucleotide having a target binding region and a second polynucleotide bound to the labeled probe in a second polynucleotide binding region thereof. The second binding region is at least partially coextensive with the target binding region. This reagent complex is contacted with a sample under conditions in which the target sequence binds to the labeled probe, displacing the second polynucleotide. Labeled probe/target hybrids are separated from intact reagent complexes and the presence and amount of these hybrids are determined.

    摘要翻译： 提供了用于确定生物样品中预定靶核苷酸序列的存在和量的方法。 该方法使用由具有靶结合区域的标记的探针多核苷酸和在其第二多核苷酸结合区域中与标记探针结合的第二多核苷酸组成的试剂复合物。 第二结合区域与靶结合区域至少部分共延伸。 在靶序列与标记的探针结合的条件下使该试剂复合物与样品接触，置换第二多核苷酸。 将标记的探针/靶杂交体与完整的试剂配合物分离，并确定这些杂交体的存在和量。