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31. 发明授权

US5658738A Bi-directional oligonucleotides that bind thrombin 失效
标题翻译：结合凝血酶的双向寡核苷酸
公开(公告)号：US5658738A
公开(公告)日：1997-08-19
申请号：US539516
申请日：1995-12-11
申请人： James G. Nadeau , Mary Lee Ciolkowski , Erwin A. Vogler
发明人： James G. Nadeau , Mary Lee Ciolkowski , Erwin A. Vogler
IPC分类号： A61K38/00 , C07H21/00 , C12N15/113 , C12Q1/68 , A01N43/72 , C07H21/04
CPC分类号： C12N15/1137 , C07H21/00 , C12Q1/6811 , A61K38/00 , C12N2310/13 , C12N2310/317 , C12N2310/3183
摘要： The present invention relates to bi-directional nucleic acid ligand compounds wherein at least two oligonucleotides of opposite sequence polarity are linked to a connecting compound at their same respective terminii; either the 5' terminii or the 3' terminii. These compounds are useful for binding protein or small molecule targets and thus may be used as diagnostic or therapeutic agents.
摘要翻译：本发明涉及双向核酸配体化合物，其中相对序列极性的至少两个寡核苷酸在相同的末端与连接化合物连接; 5'末端或3'末端。这些化合物可用于结合蛋白质或小分子靶标，因此可用作诊断或治疗剂。

32. 发明授权

US5561044A Detection of mycobacteria by multiplex strand displacement nucleic acid amplification 失效
标题翻译：通过多重链置换核酸扩增检测分枝杆菌
公开(公告)号：US5561044A
公开(公告)日：1996-10-01
申请号：US398305
申请日：1995-03-03
申请人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
发明人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
IPC分类号： C12N15/09 , C12Q1/04 , C12Q1/68 , C12R1/32 , C12R1/33 , C12P19/34
CPC分类号： C12Q1/689 , C12Q1/6851 , C12Q2600/16
摘要： Primers and methods for adapter-mediated multiplex amplification of the IS6110 insertion element of Mycobacterium tuberculosis (M.tb) and the 16S ribosomal gene of Mycobacterium tuberculosis, useful for simultaneously detecting and/or identifying species of the M. tuberculosis complex and other clinically relevant Mycobacterium species. Multiplex Strand Displacement Amplification (SDA) is used in a single amplification reaction which is capable of simultaneously identifying M. tuberculosis and providing a screen for substantially all of the clinically relevant species of Mycobacteria. Also disclosed are methods for adapter-mediated multiplex amplification of multiple target sequences and a single internal control sequence for determination of sample efficacy or quantitation of the targets. In a preferred embodiment, an internal control sequence is included in the amplification reaction and coamplified with the IS6110 and 16S target sequences as an indication of sample amplification activity or to quantitate the initial amount of target sequences in the sample.
摘要翻译：针对结核分枝杆菌（M.tb）的IS6110插入元件和结核分枝杆菌的16S核糖体基因的适配器介导的多重扩增的引物和方法，可用于同时检测和/或鉴定结核分枝杆菌复合物和其他临床相关的物种分枝杆菌种。多重链位移扩增（SDA）用于能够同时鉴定结核分枝杆菌并为基本上所有临床相关分枝杆菌物种提供筛选的单一扩增反应。还公开了用于多重靶序列的衔接子介导的多重扩增的方法和用于确定靶的样品功效或定量的单个内部控制序列的方法。在优选的实施方案中，内部对照序列包括在扩增反应中并与IS6110和16S靶序列共扩增，作为样品扩增活性的指示或定量样品中靶序列的初始量。

33. 发明授权

US5470723A Detection of mycobacteria by multiplex nucleic acid amplification 失效
标题翻译：通过多重核酸扩增检测分枝杆菌
公开(公告)号：US5470723A
公开(公告)日：1995-11-28
申请号：US111076
申请日：1993-08-24
申请人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
发明人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
IPC分类号： C12N15/09 , C12Q1/04 , C12Q1/68 , C12R1/32 , C12R1/33 , C07H15/12 , C07H17/00 , C12P19/34
CPC分类号： C12Q1/689 , C12Q1/6851 , C12Q2600/16
摘要： Primers and methods for adapter-mediated multiplex amplification of the IS6110 insertion element of Mycobacterium tuberculosis (M.tb) and the 16S ribosomal gene of Mycobacterium tuberculosis, useful for simultaneously detecting and/or identifying species of the M. tuberculosis complex and other clinically relevant Mycobacterium species. Multiplex Strand Displacement Amplification (SDA) is used in a single amplification reaction which is capable of simultaneously identifying M. tuberculosis and providing a screen for substantially all of the clinically relevant species of Mycobacteria. Also disclosed are methods for adapter-mediated multiplex amplification of multiple target sequences and a single internal control sequence for determination of sample efficacy or quantitation of the targets. In a preferred embodiment, an internal control sequence is included in the amplification reaction and coamplified with the IS6110 ant 16S target sequences as an indication of sample amplification activity or to quantitate the initial amount of target sequences in the sample.
摘要翻译：针对结核分枝杆菌（M.tb）的IS6110插入元件和结核分枝杆菌的16S核糖体基因的适配器介导的多重扩增的引物和方法，可用于同时检测和/或鉴定结核分枝杆菌复合物和其他临床相关的物种分枝杆菌种。多重链位移扩增（SDA）用于能够同时鉴定结核分枝杆菌并为基本上所有临床相关分枝杆菌物种提供筛选的单一扩增反应。还公开了用于多重靶序列的衔接子介导的多重扩增的方法和用于确定靶的样品功效或定量的单个内部控制序列的方法。在优选的实施方案中，扩增反应中包含内部控制序列并与IS6110蚂蚁16S靶序列共扩增，作为样品扩增活性的指示或定量样品中靶序列的初始量。

34. 发明授权

US5457027A Internal controls for isothermal nucleic acid amplification reactions 失效
标题翻译：用于等温核酸扩增反应的内部对照
公开(公告)号：US5457027A
公开(公告)日：1995-10-10
申请号：US058648
申请日：1993-05-05
申请人： James G. Nadeau , Michael C. Little
发明人： James G. Nadeau , Michael C. Little
IPC分类号： C12Q1/68 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6865 , C12Q1/6851 , C12Q1/689
摘要： Methods employing internal oligonucleotide standards in isothermal nucleic acid amplification reactions to determine the efficacy of the amplification reaction and to quantify pre-amplification target levels.
摘要翻译：在等温核酸扩增反应中使用内部寡核苷酸标准物来确定扩增反应的功效并量化预扩增目标水平的方法。

35. 发明授权

US09404160B2 Methods for the detection of microorganisms 有权
标题翻译：微生物检测方法
公开(公告)号：US09404160B2
公开(公告)日：2016-08-02
申请号：US13518211
申请日：2010-12-21
申请人： James G. Nadeau , Bernard J. H. Verwer
发明人： James G. Nadeau , Bernard J. H. Verwer
IPC分类号： C12Q1/68
CPC分类号： C12Q1/689 , C12Q2600/16
摘要： Presented herein are methods for the detection of the presence or absence of one or more microorganisms in a sample. The method deploys a plurality of probe sets to detect a plurality of microorganisms. The probes in the probe set are detectably labeled. At least one probe set has probes labeled with a combination of detectable labels. The number of detectable labels used in the plurality of probe sets numbers less than the number of microorganisms being detected by the probe set.
摘要翻译：本文提出的是用于检测样品中一种或多种微生物的存在或不存在的方法。该方法部署多个探针组以检测多个微生物。探针组中的探针可以被可检测地标记。至少一个探针组具有用可检测标记的组合标记的探针。在多个探针组中使用的可检测标记的数量小于由探针组检测到的微生物数量。

36. 发明申请

US20120329050A1 METHODS FOR THE DETECTION OF MICROORGANISMS 有权
标题翻译：检测微生物的方法
公开(公告)号：US20120329050A1
公开(公告)日：2012-12-27
申请号：US13518211
申请日：2010-12-21
申请人： James G. Nadeau , Bernard J.H. Verwer
发明人： James G. Nadeau , Bernard J.H. Verwer
IPC分类号： C12Q1/68 , G01N21/76 , C12M1/34 , G01N21/64
CPC分类号： C12Q1/689 , C12Q2600/16
摘要： Presented herein are methods for the detection of the presence or absence of one or more microorganisms in a sample. The method deploys a plurality of probe sets to detect a plurality of microorganisms. The probes in the probe set are detectably labeled. At least one probe set has probes labeled with a combination of detectable labels. The number of detectable labels used in the plurality of probe sets numbers less than the number of microorganisms being detected by the probe set.
摘要翻译：本文提出的是用于检测样品中一种或多种微生物的存在或不存在的方法。该方法部署多个探针组以检测多个微生物。探针组中的探针可以被可检测地标记。至少一个探针组具有用可检测标记的组合标记的探针。在多个探针组中使用的可检测标记的数量小于由探针组检测到的微生物数量。

37. 发明申请

US20120276538A1 SEQUENCE-SPECIFIC METHODS FOR HOMOGENEOUS, REAL-TIME DETECTION OF LAMP PRODUCTS 有权
标题翻译：序列特异性方法，用于均质，实时检测灯产品
公开(公告)号：US20120276538A1
公开(公告)日：2012-11-01
申请号：US13505598
申请日：2010-11-04
申请人： James G. Nadeau
发明人： James G. Nadeau
IPC分类号： C12Q1/68 , G01N21/65 , G01N21/64
CPC分类号： C12Q1/6865 , C12Q2525/307
摘要： Presented herein are methods and compositions for generating sequence-specific, secondary amplification products during Loop-mediated Isothermal Amplification (LAMP). Conventional LAMP produces a preponderance of high molecular weight DNA structures concatenated into self-complementary hairpins, which are not amenable to detection by routine probe-based hybridization methods, making multiplex detection of two or more targets or sequence variants in closed-tube formats extremely difficult. Provided herein, for example, are methods for generating secondary LAMP products bearing a fragment of the original target sequence embedded within low-molecular weight products that are devoid of competitive hairpin structures, the lack of which enhances probe-based detection of target sequences. These secondary products can, for example, be produced in real-time, during the LAMP process, and can provide the option of detecting multiple target sequences within a single tube using, e.g., a homogenous, real-time fluorescence format.
摘要翻译：本文提出了在环介导的等温扩增（LAMP）期间产生序列特异性二级扩增产物的方法和组合物。常规的LAMP产生连接到自身互补发夹中的高分子量DNA结构的优势，其不适于通过常规基于探针的杂交方法的检测，使得封闭管形式中的两个或更多个目标或序列变体的多重检测非常困难。例如，本文提供的是产生具有嵌入低分子量产物中的原始靶序列的片段的次级LAMP产物的方法，所述片段不含有竞争力的发夹结构，其缺乏可增强靶序列的基于探针的检测。这些次要产物可以例如在LAMP过程期间实时产生，并且可以提供使用例如均匀的实时荧光格式在单个管内检测多个靶序列的选项。

38. 发明授权

US06379888B1 Universal probes and methods for detection of nucleic acids 失效
标题翻译：用于检测核酸的通用探针和方法
公开(公告)号：US06379888B1
公开(公告)日：2002-04-30
申请号：US09406074
申请日：1999-09-27
申请人： James G. Nadeau , C. Preston Linn , J. Bruce Pitner , Cheryl H. Dean , G. Terrance Walker
发明人： James G. Nadeau , C. Preston Linn , J. Bruce Pitner , Cheryl H. Dean , G. Terrance Walker
IPC分类号： C12Q168
CPC分类号： C12Q1/6818 , Y10T436/143333 , C12Q2563/137 , C12Q2537/1373
摘要： Signal primers are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The signal primer comprises a first and a second oligonucleotide and is partially single-stranded and partially double-stranded. In the presence of target, the second oligonucleotide of the signal primer is displaced from the first and a conformational change in a reporter probe occurs which changes the distance between the members of a donor/quencher dye pair linked to the reporter probe. The change in proximity between the dyes causes an increase or a decrease in fluorescence quenching, which is detected as an indication of the presence of the target sequence.
摘要翻译：信号引物用于通过荧光猝灭机制检测核酸靶序列。信号引物包含第一和第二寡核苷酸并且是部分单链和部分双链的。在靶的存在下，信号引物的第二寡核苷酸从第一个位点移位，并且发生报告物探针中的构象变化，其改变与报道探针连接的供体/猝灭剂染料对的成员之间的距离。染料之间接近度的变化导致荧光淬灭的增加或减少，其被检测为靶序列存在的指示。

39. 发明授权

US06261784B1 Detection of nucleic acids by strand displacement 有权
标题翻译：通过链置换检测核酸
公开(公告)号：US06261784B1
公开(公告)日：2001-07-17
申请号：US09599164
申请日：2000-06-22
申请人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
发明人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
IPC分类号： C12Q168
CPC分类号： C12Q1/6818 , C12Q2537/1373 , C12Q2531/143 , C12Q2531/119
摘要： Detector nucleic acids are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The detector nucleic acid comprises at least two oligonucleotides and is partially single-stranded and partially double-stranded. One of the two dyes of a donor/acceptor dye pair is linked to the first oligonucleotide and the other is linked to a second oligonucleotide such that they are in close spatial proximity when the first and second oligonucleotides are base-paired and donor fluorescence is quenched. A single second oligonucleotide may be hybridized to the first oligonucleotide or multiple second oligonucleotides may be hybridized to the first oligonucleotide and to each other, forming a junction structure comprising multiple donor/acceptor dye pairs. The detector oligonucleotide retains its partially single-stranded and partially double-stranded conformation in the absence of target. In the presence of target, however, the second oligonucleotide(s) of the detector nucleic acid is/are completely or partially displaced from the first, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence which may be detected as an indication of the presence of the target sequence.
摘要翻译：检测器核酸用于通过荧光猝灭机制检测核酸靶序列。检测器核酸包含至少两个寡核苷酸并且是部分单链和部分双链的。供体/受体染料对的两种染料之一与第一寡核苷酸连接，另一种与第二寡核苷酸连接，使得当第一和第二寡核苷酸是碱基配对并且供体荧光淬灭时它们处于紧密的空间接近。单个第二寡核苷酸可以与第一寡核苷酸杂交，或者多个第二寡核苷酸可以与第一寡核苷酸和彼此杂交，形成包含多个供体/受体染料对的连接结构。检测器寡核苷酸在不存在目标时保留其部分单链和部分双链构象。然而，在靶存在下，检测器核酸的第二寡核苷酸与第一寡核苷酸完全或部分位移，从而增加供体和受体染料之间的距离并引起荧光的变化，其可被检测为指示靶序列的存在。

40. 发明授权

US6130047A Detection of nucleic acids by fluorescence quenching 失效
标题翻译：通过荧光猝灭检测核酸
公开(公告)号：US6130047A
公开(公告)日：2000-10-10
申请号：US235583
申请日：1999-01-22
申请人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
发明人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
IPC分类号： G01N21/64 , C07H21/00 , C12N15/09 , C12Q1/68 , G01N33/542 , G01N33/566 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6818
摘要： Detector nucleic acids are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The detector nucleic acid comprises at least two oligonucleotides and is partially single-stranded and partially double-stranded. One of the two dyes of a donor/acceptor dye pair is linked to the first oligonucleotide and the other is linked to a second oligonucleotide such that they are in close spatial proximity when the first and second oligonucleotides are base-paired and donor fluorescence is quenched. A single second oligonucleotide may be hybridized to the first oligonucleotide or multiple second oligonucleotides may be hybridized to the first oligonucleotide and to each other, forming a junction structure comprising multiple donor/acceptor dye pairs. The detector oligonucleotide retains its partially single-stranded and partially double-stranded conformation in the absence of target. In the presence of target, however, the second oligonucleotide(s) of the detector nucleic acid is/are completely or partially displaced from the first, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence which may be detected as an indication of the presence of the target sequence.
摘要翻译：检测器核酸用于通过荧光猝灭机制检测核酸靶序列。检测器核酸包含至少两个寡核苷酸并且是部分单链和部分双链的。供体/受体染料对的两种染料之一与第一寡核苷酸连接，另一种与第二寡核苷酸连接，使得当第一和第二寡核苷酸是碱基配对并且供体荧光淬灭时它们处于紧密的空间接近。单个第二寡核苷酸可以与第一寡核苷酸杂交，或者多个第二寡核苷酸可以与第一寡核苷酸和彼此杂交，形成包含多个供体/受体染料对的连接结构。检测器寡核苷酸在不存在目标时保留其部分单链和部分双链构象。然而，在靶存在下，检测器核酸的第二寡核苷酸与第一寡核苷酸完全或部分位移，从而增加供体和受体染料之间的距离并引起荧光的变化，其可被检测为指示靶序列的存在。

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