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21.发明授权
公开(公告)号：US5595891A
公开(公告)日：1997-01-21
申请号：US555323
申请日：1990-07-19
申请人： Samuel Rose , Linda M. Western , Martin Becker , Edwin F. Ullman
发明人： Samuel Rose , Linda M. Western , Martin Becker , Edwin F. Ullman
CPC分类号： C12Q1/6853 , C12N15/10 , C12Q1/686
摘要： A method is disclosed for producing a single stranded polydeoxynucleotide having two segments that are non-contiguous and complementary with each other. The method comprises the steps of providing in combination (1) a polynucleotide having two non-contiguous, non-complementary nucleotide sequences S1 and S2 wherein S2 is 5' of S1 and is at least ten deoxynucleotides long and (2) an extender probe comprised of two deoxynucleotide sequences, wherein the sequence at the 3'-end of the extender probe is hybridizable with S1 and the other of the deoxynucleotide sequences is homologous to S2 and (b) extending the extender probe along the polynucleotide. The method can also comprise providing in the combination a polydoxynucleotide primer capable of hybridizing at least at its 3'-end with a nucleotide sequence complementary to S2 under conditions where (1) the extended extender probe is rendered single stranded, (2) the polydeoxynucleotide primer hybridizes with and is extended along the extended extender probe to form a duplex comprising extended primer, (3) the extended primer is dissociated from the duplex, and (4) the primer hybridizes with and is extended along the extended primer to form a duplex comprising extended primer, and repeating steps (3) and (4). The method finds particular application in the detection of polynucleotide analytes.
摘要翻译： 公开了一种用于生产具有彼此不连续和互补的两个区段的单链多脱氧核苷酸的方法。 该方法包括以下步骤：组合（1）具有两个非连续的非互补核苷酸序列S1和S2的多核苷酸，其中S2是S1的5'，长至少十个脱氧核苷酸，和（2） 的两个脱氧核苷酸序列，其中扩增物探针的3'末端的序列可与S1杂交，而另一个脱氧核苷酸序列与S2同源，并且（b）沿多核苷酸扩展扩增体探针。 该方法还可以包括提供能够在其3'末端与至少与S2互补的核苷酸序列的多核苷酸引物提供，其中（1）扩展扩增物探针是单链的，（2）多脱氧核苷酸 引物与扩展的扩增物探针杂交并延伸，以形成包含延伸引物的双链体，（3）延伸引物与双链体解离，和（4）引物与延伸引物杂交并延伸以形成双链体 包括延伸引物，并重复步骤（3）和（4）。 该方法在多核苷酸分析物的检测中具有特殊应用。
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公开(公告)号：US5536644A
公开(公告)日：1996-07-16
申请号：US353481
申请日：1994-12-09
IPC分类号： B03C1/01 , C12Q1/24 , G01N33/538 , G01N33/543 , G01N33/555
CPC分类号： C12Q1/24 , B03C1/01 , G01N33/538 , G01N33/54333 , Y10S436/829
摘要： A method is disclosed for separating a substance from a liquid medium. The method comprises combining the liquid medium containing the substance with magnetic particles under conditions for non-specific chemical binding of the magnetic particles. Thereafter, the medium is subjected to a magnetic field gradient to separate the particles from the medium. The preferred non-specific binding is achieved as the result of charge interactions between the particles usually by means of a polyionic reagent. The method of the invention has particular application to the separation of cells and microorganisms from aqueous suspensions and also to the determination of an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp). The sample is combined in an assay medium with magnetic particles and a sbp member complementary to the analyte. Magnetic or non-magnetic particles capable of specific binding to the analyte or its complementary sbp member must be included in the assay medium. The combination is made under conditions for non-specifically aggregating the magnetic particles or coaggregating the magnetic and non-magnetic particles when non-magnetic particles are present. The assay medium is subjected to a magnetic field gradient to separate the aggregated particles from the medium. Then, the medium or the particles are examined for the presence or amount of the analyte or an sbp member, the binding of which is affected by the presence of the analyte.
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公开(公告)号：US5342759A
公开(公告)日：1994-08-30
申请号：US333745
申请日：1989-04-03
申请人： David J. Litman , Edwin F. Ullman
发明人： David J. Litman , Edwin F. Ullman
IPC分类号： C12N11/00 , C12Q1/26 , G01N33/52 , G01N33/542 , G01N33/543 , G01N33/58
CPC分类号： G01N33/543 , G01N33/52 , G01N33/542 , G01N33/54306 , G01N33/581 , Y10S435/805 , Y10S435/81 , Y10S435/967 , Y10S435/97 , Y10S435/973 , Y10S435/975 , Y10S436/80 , Y10S436/805 , Y10S436/806 , Y10S436/808 , Y10S436/811 , Y10S436/815 , Y10S436/82 , Y10S436/826
摘要： An assay method and compositions are provided for determining the presence of an analyte in a sample. The analyte is a member of an immunological pair (mip) comprising ligand and receptor. By providing a first measurement surface capable of specifically binding a labelled reagent in an amount depending upon the presence of analyte in the sample and a second calibration surface capable of binding a second labeled reagent in a manner unaffected by the presence of analyte in the sample, calibration of individual tests can be accomplished simulataneously with the performance of the test itself. A signal producing system includes an enzyme bonded to a mip which defines the first labeled reagent for binding to the measurement surface and the same enzyme conjugated to a ligand capable of binding to the calibration surface. Preferably, both labeled reagents have the same composition and the calibration surface includes anti-(first enzyme).
摘要翻译： 提供测定方法和组合物以确定样品中分析物的存在。 分析物是包含配体和受体的免疫对（mip）的成员。 通过提供能够以取决于样品中分析物的存在的量特异性结合标记试剂的第一测量表面和能够以不受样品中分析物的存在影响的方式结合第二标记试剂的第二校准表面， 单独测试的校准可以与测试本身的性能同时完成。 信号产生系统包括键合到mip的酶，其限定用于结合测量表面的第一标记试剂和与能够结合校准表面的配体缀合的相同酶。 优选地，两种标记的试剂具有相同的组成，并且校准表面包括抗（第一酶）。
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公开(公告)号：US5332662A
公开(公告)日：1994-07-26
申请号：US923080
申请日：1992-07-31
申请人： Edwin F. Ullman
发明人： Edwin F. Ullman
IPC分类号： C07D219/06 , C12Q1/28 , G01N33/536 , G01N33/542
CPC分类号： C12Q1/28 , C07D219/06 , C12Q2326/00 , Y10S435/81
摘要： Methods and compositions are disclosed for determining a peroxidatively active substance (PAS). The methods comprise the step of detecting a fluorescent signal produced upon cleavage of a compound of the formula F-L-Q, wherein F is a fluorester capable of producing the signal, Q is a quencher capable of quenching the signal when linked to F, and L is a bond, or a linking group having a bond, wherein the bond is capable of being cleaved by a reaction of the PAS with a substrate of the PAS and a hydrogen donor wherein the cleavage of the bond substantially reduces the quenching. The methods have application in a wide variety of systems including assays and improved assays for analytes. Also disclosed are kits for conducting the methods and improvements in accordance with the present invention.
摘要翻译： 公开了用于测定过氧化物活性物质（PAS）的方法和组合物。 所述方法包括检测在切割式FLQ化合物时产生的荧光信号的步骤，其中F是能够产生该信号的荧光素，Q是能够在与F连接时淬灭该信号的猝灭剂，L是 键或具有键的连接基团，其中该键能够通过PAS与PAS的底物和氢供体的反应而被切割，其中键的裂解基本上减少了淬灭。 该方法可应用于各种系统，包括分析物的分析和改进的测定。 还公开了用于进行根据本发明的方法和改进的套件。
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公开(公告)号：US5256575A
公开(公告)日：1993-10-26
申请号：US755203
申请日：1991-09-04
申请人： Mae W. Hu , Kirk Schulkamp , Cheng-I Lin , Edwin F. Ullman
发明人： Mae W. Hu , Kirk Schulkamp , Cheng-I Lin , Edwin F. Ullman
IPC分类号： C07C235/34 , C07C235/52 , C07C259/10 , C07C309/15 , G01N33/53 , G01N33/78 , G01N33/94 , G01N33/566
CPC分类号： C07C309/15 , C07C235/34 , C07C235/52 , C07C259/10 , G01N33/5306 , G01N33/78 , G01N33/94 , Y10S435/962 , Y10S435/975 , Y10S436/825
摘要： A kit for inactivating interfering binding proteins in a immunoassay for a member of a specific binding pair (sbp). The method comprises including in an assay medium containing a sample suspected of containing an sbp member and an interfering binding protein an effective amount of a water soluble compound having two substituted or unsubstituted phenyl groups linked to a common atom. When the sbp member or its sbp partner has two phenyl groups linked to a common atom, the compound has a number of-groups other than hydrogen attached to the phenyl groups and the atom that differs by at least two from the number of such groups on the sbp member. When the sbp member or its sbp partner has two phenyl groups linked to a common atom and the binding protein is not an antibody, the compound has only one group other than hydrogen attached to a phenyl group or the common atom. The methods have particular application in avoiding-cross-reactivity of non-analyte materials in a sample with immunachemical reagents used in such assay. The methods-have application also in disrupting complexer between an analyte to be determined and other materials to that one can accurately determine the amount of an analyte in a sample.
摘要翻译： 用于在特异性结合对（sbp）的成员的免疫测定中灭活干扰结合蛋白的试剂盒。 该方法包括在含有疑似含有sbp成员和干扰结合蛋白的样品的测定培养基中包含有效量的具有两个与常见原子连接的取代或未取代的苯基的水溶性化合物。 当sbp成员或其sbp伴侣具有连接到共同原子的两个苯基时，该化合物具有与苯基连接的氢以外的多个基团，并且与该基团的数目不同的至少两个的原子在 sbp成员。 当sbp成员或其sbp伴侣具有连接到共同原子的两个苯基并且结合蛋白不是抗体时，该化合物除了连接到苯基或共同原子上的氢以外仅具有一个基团。 该方法特别适用于避免样品中非分析物质与这种测定中使用的免疫化学试剂的交叉反应。 所述方法还应用于破坏要测定的待分析物与其他材料之间的复合物，以使得可以准确地确定样品中分析物的量。
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公开(公告)号：US5136095A
公开(公告)日：1992-08-04
申请号：US278870
申请日：1988-12-01
IPC分类号： G01N33/543
CPC分类号： G01N33/54333 , G01N33/54313 , G01N33/54393 , Y10S436/808 , Y10S436/829
摘要： Compounds and methods are disclosed for reversibly aggregating particles suspended in a liquid medium. The method comprises combining the liquid medium containing the particles with a polyionic polymer capable of aggregating the particles under conditions suitable for such aggregation. Thereafter, the particles are contacted with a chemical reagent capable of cleaving the polyionic polymer under conditions sufficient to reverse the aggregation. Optionally, magnetic particles are added to the liquid medium in the present method under conditions for non-specific binding and the medium including the aggregates is subjected to a magnetic field gradient to separate the aggregates from the medium. The compounds of the present invention are polyions. The aggregation of the particles is reversible upon contact with chemical agents which cleave at least some of the bonds within the polyionic polymer.
摘要翻译： 公开了用于可逆地聚集悬浮在液体介质中的颗粒的化合物和方法。 该方法包括将含有颗粒的液体介质与能够在适于这种聚集的条件下聚集颗粒的聚离子聚合物组合。 此后，使颗粒与能够在足以反向聚集的条件下裂解聚离子聚合物的化学试剂接触。 任选地，在本方法中，在非特异性结合的条件下，将磁性颗粒加入到液体介质中，并且将包含聚集体的介质进行磁场梯度以将聚集体与介质分离。 本发明的化合物是聚阴离子。 当与化学试剂接触时，颗粒的聚集是可逆的，其切割聚离子聚合物内的至少一些键。
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公开(公告)号：US5104812A
公开(公告)日：1992-04-14
申请号：US441688
申请日：1989-11-27
申请人： Nurith Kurn , Rajesh D. Patel , Martin Becker , Edwin F. Ullman
发明人： Nurith Kurn , Rajesh D. Patel , Martin Becker , Edwin F. Ullman
IPC分类号： G01N33/52 , G01N33/558
CPC分类号： G01N33/558 , G01N33/521 , Y10S435/805 , Y10S435/81 , Y10S436/807 , Y10T436/2575
摘要： Disclosed are devices and methods for interrupting capillary flow of a liquid between two pieces of bibulous material which, prior to actuation, are in a capillary flow relationship to each other. In particular, the capillary flow relationship of two pieces of bibulous material is interrupted by utilizing a liquid expandable piece of bibulous material.
摘要翻译： 公开了用于中断两片吸水材料之间的液体的毛细管流动的装置和方法，其在致动之前彼此处于毛细管流动关系中。 特别地，通过利用吸水材料的液体可膨胀片，中断两片吸水材料的毛细管流动关系。
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公开(公告)号：US5076950A
公开(公告)日：1991-12-31
申请号：US533622
申请日：1990-06-05
IPC分类号： B03C1/01 , C12Q1/24 , G01N33/538 , G01N33/543
CPC分类号： C12Q1/24 , B03C1/01 , G01N33/538 , G01N33/54333 , Y10S436/829
摘要： A method is disclosed for separating a substance from a liquid medium. The method comprises combining the liquid medium containing the substance with magnetic particles under conditions for non-specific chemical binding of the magnetic particles. Thereafter, the medium is subjected to a magnetic field gradient to separate the particles from the medium. The preferred non-specific binding is achieved as the result of charge interactions between the particles usually by means of a polyionic reagent. The method of the invention has particular application to the separation of cells and microorganisms from aqueous suspensions and also to the determination of an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp). The sample is combined in an assay medium with magnetic particles and a sbp member complementary to the analyte. Magnetic or non-magnetic particles capable of specific binding to the analyte or its complementary sbp member must be included in the assay medium. The combination is made under conditions for non-specifically aggregating the magnetic particles or coaggregating the magnetic and non-magnetic particles when non-magnetic particles are present. The assay medium is subjected to a magnetic field gradient to separate the aggregated particles from the medium. Then, the medium or the particles are examined for the presence or amount of the analyte or an sbp member, the binding of which is affected by the presence of the analyte.
摘要翻译： 公开了一种用于从液体介质中分离物质的方法。 该方法包括在含有磁性颗粒的非特异性化学结合的条件下将含有物质的液体介质与磁性颗粒组合。 此后，对介质进行磁场梯度以将颗粒与介质分离。 优选的非特异性结合是通过颗粒之间的电荷相互作用的结果实现的，通常通过聚离子试剂。 本发明的方法特别适用于从水性悬浮液中分离细胞和微生物，以及用于测定怀疑含有分析物的样品中的分析物。 分析物是特异性结合对（sbp）的成员。 将样品在测定介质中与磁性颗粒和与分析物互补的sbp构件组合。 能够与分析物或其互补sbp成员特异性结合的磁性或非磁性粒子必须包含在测定介质中。 在非磁性颗粒存在的情况下，在非特异性聚集磁性颗粒或使磁性和非磁性颗粒共聚的条件下进行组合。 对测定介质进行磁场梯度以将聚集的颗粒与介质分离。 然后，检查介质或颗粒的分析物或sbp构件的存在或量，其结合受分析物的存在的影响。
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公开(公告)号：US4935147A
公开(公告)日：1990-06-19
申请号：US262771
申请日：1988-10-26
IPC分类号： B03C1/01 , C12Q1/24 , G01N33/538 , G01N33/543
CPC分类号： B03C1/01 , C12Q1/24 , G01N33/538 , G01N33/54333
摘要： A method is disclosed for separating a substance from a liquid medium. The method comprises combining the liquid medium containing the substance with magnetic particles under conditions for non-specific chemical binding of the magnetic particles. Thereafter, the medium is subjected to a magnetic field gradient to separate the particles from the medium. The preferred non-specific binding is achieved as the result of charge interactions between the particles usually by means of a polyionic reagent. The method of the invention has particular application to the separation of cells and microorganisms from aqueous suspensions and also to the determination of an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp). The sample is combined in an assay medium with magnetic particles and a sbp member complementary to the analyte. Magnetic or non-magnetic particles capable of specific binding to the analyte or its complementary sbp member must be included in the assay medium. The combination is made under conditions for non-specifically aggregating the magnetic particles or coaggregating the magnetic and non-magnetic particles when non-magnetic particles are present. The assay medium is subjected to a magnetic field gradient to separate the aggregated particles from the medium. Then, the medium or the particles are examined for the presence or amount of the analyte or an sbp member, the binding of which is affected by the presence of the analyte.
摘要翻译： 公开了一种用于从液体介质中分离物质的方法。 该方法包括在含有磁性颗粒的非特异性化学结合的条件下将含有物质的液体介质与磁性颗粒组合。 此后，对介质进行磁场梯度以将颗粒与介质分离。 优选的非特异性结合是通过颗粒之间的电荷相互作用的结果实现的，通常通过聚离子试剂。 本发明的方法特别适用于从水性悬浮液中分离细胞和微生物，以及用于测定怀疑含有分析物的样品中的分析物。 分析物是特异性结合对（sbp）的成员。 将样品在测定介质中与磁性颗粒和与分析物互补的sbp构件组合。 能够与分析物或其互补sbp成员特异性结合的磁性或非磁性粒子必须包含在测定介质中。 在非磁性颗粒存在的情况下，在非特异性聚集磁性颗粒或使磁性和非磁性颗粒共聚的条件下进行组合。 对测定介质进行磁场梯度以将聚集的颗粒与介质分离。 然后，检查介质或颗粒的分析物或sbp构件的存在或量，其结合受分析物的存在的影响。
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公开(公告)号：US4748129A
公开(公告)日：1988-05-31
申请号：US645458
申请日：1984-08-28
IPC分类号： G01N33/52 , A61K39/00 , G01N33/53 , G01N33/533 , G01N33/536 , G01N33/542 , G01N33/554 , G01N33/555 , G01N33/58 , G01N33/80
CPC分类号： G01N33/555 , G01N33/533 , G01N33/542 , G01N33/80 , Y10S436/80 , Y10S436/805 , Y10S436/807 , Y10S436/829
摘要： A method is provided for determining the presence in a sample of a member of a specific binding pair ("sbp member") consisting of ligand and its homologous receptor. The sample is combined in an aqueous medium with (1) a complementary sbp member wherein at least the sbp member or the complementary sbp member is bound to the surface of a cell and (2) a fuorescent agent capable of being incorporated into the cell. The presence of the sbp member is indicated by a change in fluorescence of the unseparated cell suspension as a result of agglutination of the cells.The present invention has particular application to blood typing, for example, for the determination of the presence of blood group antigens A, B, AB, O, and D (Rh.sub.o) and antibodies to such antigens.
摘要翻译： 提供了一种用于确定由配体及其同源受体组成的特异性结合对（“sbp成员”）的成员的样品中存在的方法。 样品在水性介质中与（1）互补sbp构件组合，其中至少sbp构件或互补sbp构件与细胞表面结合，和（2）能够结合到细胞中的荧光剂。 由于细胞凝集，未分离的细胞悬浮液的荧光变化表示sbp成员的存在。 本发明特别适用于血型分型，例如用于测定血型抗原A，B，AB，O和D（Rho）及其抗体的存在。
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