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公开(公告)号：US20080213767A1
公开(公告)日：2008-09-04
申请号：US11869054
申请日：2007-10-09
IPC分类号： C12Q1/68
CPC分类号： C12Q1/689 , C12Q1/682 , C12Q1/6823 , C12Q2521/319 , C12Q2537/149 , C12Q2521/301 , C12Q2563/101
摘要： A method is disclosed for modifying an oligonucleotide, which method has application to the detection of a polynucleotide analyte. An oligonucleotide is reversibly hybridized with a polynucleotide, for example, a polynucleotide analyte, in the presence of a 5′-nuclease under isothermal conditions. The polynucleotide analyte serves as a recognition element to enable a 5′-nuclease to cleave the oligonucleotide to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide analyte and (ii) a second fragment that lies 3′ of the first fragment (in the intact oligonucleotide) and is substantially hybridizable to the polynucleotide analyte. At least a 100-fold molar excess of the first fragment and/or the second fragment are obtained relative to the molar amount of the polynucleotide analyte. The presence of the first fragment and/or the second fragment is detected, the presence thereof indicating the presence of the polynucleotide analyte. The method has particular application to the detection of a polynucleotide analyte such as DNA. Kits for conducting methods in accordance with the present invention are also disclosed.
摘要翻译： 公开了一种修饰寡核苷酸的方法，该方法可用于检测多核苷酸分析物。 在等温条件下，在5'-核酸酶的存在下，寡核苷酸与多核苷酸例如多核苷酸分析物可逆地杂交。 多核苷酸分析物用作识别元件，使得5'-核酸酶能够切割寡核苷酸，以提供（i）与多核苷酸分析物基本上不可杂交的第一片段和（ii）位于3'末端的第3个片段 第一片段（在完整寡核苷酸中）并且与多核苷酸分析物基本上可杂交。 相对于多核苷酸分析物的摩尔量，获得至少100倍摩尔过量的第一片段和/或第二片段。 检测到第一片段和/或第二片段的存在，其存在表明多核苷酸分析物的存在。 该方法特别适用于多核苷酸分析物如DNA的检测。 还公开了用于根据本发明的方法的套件。
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公开(公告)号：US6121001A
公开(公告)日：2000-09-19
申请号：US440363
申请日：1999-11-15
CPC分类号： C12Q1/689 , C12Q1/682 , C12Q1/6823
摘要： A method is disclosed for modifying an oligonucleotide, which method has application to the detection of a polynucleotide analyte. An oligonucleotide is reversibly hybridized with a polynucleotide, for example, a polynucleotide analyte, in the presence of a 5'-nuclease under isothermal conditions. The polynucleotide analyte serves as a recognition element to enable a 5'-nuclease to cleave the oligonucleotide to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide analyte and (ii) a second fragment that lies 3' of the first fragment (in the intact oligonucleotide) and is substantially hybridizable to the polynucleotide analyte. At least a 100-fold molar excess of the first fragment and/or the second fragment are obtained relative to the molar amount of the polynucleotide analyte. The presence of the first fragment and/or the second fragment is detected, the presence thereof indicating the presence of the polynucleotide analyte. The method has particular application to the detection of a polynucleotide analyte such as DNA. Kits for conducting methods in accordance with the present invention are also disclosed.
摘要翻译： 公开了一种修饰寡核苷酸的方法，该方法可用于检测多核苷酸分析物。 在等温条件下，在5'-核酸酶的存在下，寡核苷酸与多核苷酸例如多核苷酸分析物可逆地杂交。 多核苷酸分析物用作识别元件，使得5'-核酸酶能够切割寡核苷酸，以提供（i）与多核苷酸分析物基本上不可杂交的第一片段和（ii）位于3'末端的第3个片段 第一片段（在完整寡核苷酸中）并且与多核苷酸分析物基本上可杂交。 相对于多核苷酸分析物的摩尔量，获得至少100倍摩尔过量的第一片段和/或第二片段。 检测到第一片段和/或第二片段的存在，其存在表明多核苷酸分析物的存在。 该方法特别适用于多核苷酸分析物如DNA的检测。 还公开了用于根据本发明的方法的套件。
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3.发明授权Oligonucleotide modification, signal amplification, and nucleic acid detection by target-catalyzed product formation 失效通过靶催化产物形成的寡核苷酸修饰，信号扩增和核酸检测
公开(公告)号：US6110677A
公开(公告)日：2000-08-29
申请号：US15949
申请日：1998-01-30
CPC分类号： C12Q1/689 , C12Q1/682 , C12Q1/6823
摘要： A method is disclosed for modifying an oligonucleotide, which method has application to the detection of a polynucleotide analyte. An oligonucleotide is reversibly hybridized with a polynucleotide, for example, a polynucleotide analyte, in the presence of a 5'-nuclease under isothermal conditions. The polynucleotide analyte serves as a recognition element to enable a 5'-nuclease to cleave the oligonucleotide to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide analyte and (ii) a second fragment that lies 3' of the first fragment (in the intact oligonucleotide) and is substantially hybridizable to the polynucleotide analyte. At least a 100-fold molar excess of the first fragment and/or the second fragment are obtained relative to the molar amount of the polynucleotide analyte. The presence of the first fragment and/or the second fragment is detected, the presence thereof indicating the presence of the polynucleotide analyte. The method has particular application to the detection of a polynucleotide analyte such as DNA. Kits for conducting methods in accordance with the present invention are also disclosed.
摘要翻译： 公开了一种修饰寡核苷酸的方法，该方法可用于检测多核苷酸分析物。 在等温条件下，在5'-核酸酶的存在下，寡核苷酸与多核苷酸例如多核苷酸分析物可逆地杂交。 多核苷酸分析物用作识别元件，使得5'-核酸酶能够切割寡核苷酸，以提供（i）与多核苷酸分析物基本上不可杂交的第一片段和（ii）位于3'末端的第3个片段 第一片段（在完整寡核苷酸中）并且与多核苷酸分析物基本上可杂交。 相对于多核苷酸分析物的摩尔量，获得至少100倍摩尔过量的第一片段和/或第二片段。 检测到第一片段和/或第二片段的存在，其存在表明多核苷酸分析物的存在。 该方法特别适用于多核苷酸分析物如DNA的检测。 还公开了用于根据本发明的方法的套件。
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公开(公告)号：US5882857A
公开(公告)日：1999-03-16
申请号：US475283
申请日：1995-06-07
CPC分类号： C12Q1/6848
摘要： The present invention relates to an improvement in a method for amplifying a target sequence of a target polynucleotide. The method comprises combining a sample suspected of containing the target polynucleotide with reagents for amplifying the target sequence and subjecting the combination to conditions wherein the target sequence if present is amplified. The present improvement comprises including in the combination a control oligonucleotide and a control polynucleotide that has a sequence that is hybridizable with the control oligonucleotide. When the control oligonucleotide is bound to the control polynucleotide, the ability of a primer to chain extend along the control polynucleotide is reduced. Optionally, the control oligonucleotide is part of the control polynucleotide. The method finds particular application in the area of nucleic acid amplification and detection.
摘要翻译： 本发明涉及扩增靶多核苷酸的靶序列的方法的改进。 该方法包括将怀疑含有目标多核苷酸的样品与用于扩增靶序列的试剂结合，并使该组合进行其中如果存在的靶序列被扩增的条件。 本发明的改进包括控制寡核苷酸和具有可与对照寡核苷酸杂交的序列的对照多核苷酸的组合。 当对照寡核苷酸与对照多核苷酸结合时，引物沿着对照多核苷酸扩链的能力降低。 任选地，对照寡核苷酸是对照多核苷酸的一部分。 该方法在核酸扩增和检测领域具有特殊应用。
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公开(公告)号：US06368803B1
公开(公告)日：2002-04-09
申请号：US09608721
申请日：2000-06-30
IPC分类号： C12Q168
CPC分类号： C12Q1/689 , C12Q1/682 , C12Q1/6823 , C12Q2521/319 , C12Q2537/149 , C12Q2521/301 , C12Q2563/101
摘要： A method is disclosed for modifying an oligonucleotide, which method has application to the detection of a polynucleotide analyte. An oligonucleotide is reversibly hybridized with a polynucleotide, for example, a polynucleotide analyte, in the presence of a 5′-nuclease under isothermal conditions. The polynucleotide analyte serves as a recognition element to enable a 5′-nuclease to cleave the oligonucleotide to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide analyte and (ii) a second fragment that lies 3′ of the first fragment (in the intact oligonucleotide) and is substantially hybridizable to the polynucleotide analyte. At least a 100-fold molar excess of the first fragment and/or the second fragment are obtained relative to the molar amount of the polynucleotide analyte. The presence of the first fragment and/or the second fragment is detected, the presence thereof indicating the presence of the polynucleotide analyte. The method has particular application to the detection of a polynucleotide analyte such as DNA. Kits for conducting methods in accordance with the present invention are also disclosed.
摘要翻译： 公开了一种修饰寡核苷酸的方法，该方法可用于检测多核苷酸分析物。 在等温条件下，在5'-核酸酶的存在下，寡核苷酸与多核苷酸例如多核苷酸分析物可逆地杂交。 多核苷酸分析物用作识别元件，使得5'-核酸酶能够切割寡核苷酸，以提供（i）与多核苷酸分析物基本上不可杂交的第一片段和（ii）位于3'末端的第3个片段 第一片段（在完整寡核苷酸中）并且与多核苷酸分析物基本上可杂交。 相对于多核苷酸分析物的摩尔量，获得至少100倍摩尔过量的第一片段和/或第二片段。 检测到第一片段和/或第二片段的存在，其存在表明多核苷酸分析物的存在。 该方法特别适用于多核苷酸分析物如DNA的检测。 还公开了用于根据本发明的方法的套件。
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公开(公告)号：US5882867A
公开(公告)日：1999-03-16
申请号：US486301
申请日：1995-06-07
CPC分类号： C12Q1/6858
摘要： A method is disclosed for detecting a target polynucleotide sequence. The method comprises incubating an oligonucleotide with the target polynucleotide sequence and a nucleotide polymerase under isothermal conditions wherein at least one nucleotide is added to the 3'-terminus of the oligonucleotide to provide an extended oligonucleotide having the additional nucleotides. The presence of extended oligonucleotide is detected as an indication of the presence of the target polynucleotide sequence. The method has particular application to the detection of DNA.
摘要翻译： 公开了用于检测靶多核苷酸序列的方法。 该方法包括在等温条件下将寡核苷酸与靶多核苷酸序列和核苷酸聚合酶一起温育，其中至少一个核苷酸加入到寡核苷酸的3'末端以提供具有额外核苷酸的延伸寡核苷酸。 检测到延伸寡核苷酸的存在作为靶多核苷酸序列存在的指示。 该方法特别适用于检测DNA。
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公开(公告)号：US5792614A
公开(公告)日：1998-08-11
申请号：US691627
申请日：1996-08-02
CPC分类号： C12Q1/689 , C12Q1/682 , C12Q1/6823
摘要： A method is disclosed for modifying an oligonucleotide, which method has application to the detection of a polynucleotide analyte. An oligonucleotide is reversibly hybridized with a polynucleotide, for example, a polynucleotide analyte, in the presence of a 5'-nuclease under isothermal conditions. The polynucleotide analyte serves as a recognition element to enable a 5'-nuclease to cleave the oligonucleotide to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide analyte and (ii) a second fragment that lies 3' of the first fragment (in the intact oligonucleotide) and is substantially hybridizable to the polynucleotide analyte. At least a 100-fold molar excess of the first fragment and/or the second fragment are obtained relative to the molar amount of the polynucleotide analyte. The presence of the first fragment and/or the second fragment is detected, the presence thereof indicating the presence of the polynucleotide analyte. The method has particular application to the detection of a polynucleotide analyte such as DNA. Kits for conducting methods in accordance with the present invention are also disclosed.
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8.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：US6124090A
公开(公告)日：2000-09-26
申请号：US438149
申请日：1995-05-09
CPC分类号： C12Q1/682 , C12Q1/6844 , C12Q1/6853 , C12Q1/6862 , C12Q1/689
摘要： A method is disclosed for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte. The method comprises (a) forming as a result of the presence of an analyte a single stranded polynucleotide comprising a target polynucleotide binding sequence flanked by first and second polynucleotide sequences that differ from the sequence of the analyte or a sequence complementary to the analyte sequence, (b) forming multiple copies of the single stranded polynucleotide, and (c) detecting the single stranded polynucleotide. Also disclosed is a method of producing at least one copy of a single stranded polynucleotide. The method comprises (a) forming in the presence of nucleoside triphosphates and template dependent polynucleotide polymerase an extension of a polynucleotide primer at least the 3'-end of which has at least a 10 base sequence hybridizable with a second sequence flanking the 3'-end of the single stranded polynucleotide, the second sequence being partially or fully complementary with at least a 10 base first sequence flanking the 5' end of the single stranded polynucleotide, (b) dissociating the extended polynucleotide primer and the single stranded polynucleotide, (c) repeating step a and (d) dissociating the extended polynucleotide primer and the copy of the single stranded polynucleotide.
摘要翻译： 公开了一种用于确定疑似含有分析物的样品中多核苷酸分析物的存在的方法。 该方法包括（a）作为分析物存在的结果形成单链多核苷酸，所述单链多核苷酸包含不同于分析物序列或与分析物序列互补的序列的第一和第二多核苷酸序列侧翼的靶多核苷酸结合序列， （b）形成多重拷贝的单链多核苷酸，和（c）检测单链多核苷酸。 还公开了生产单链多核苷酸的至少一个拷贝的方法。 该方法包括（a）在核苷三磷酸和模板依赖性多核苷酸聚合酶存在下形成多核苷酸引物的延伸，所述多核苷酸引物的至少3'末端具有至少10个碱基序列，可与3'- 单链多核苷酸的末端，第二个序列与单链多核苷酸的5'端侧翼的至少10个碱基第一序列部分或完全互补，（b）解离延伸的多核苷酸引物和单链多核苷酸，（c ）重复步骤a和（d）解离延伸的多核苷酸引物和单链多核苷酸的拷贝。
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公开(公告)号：US5827649A
公开(公告)日：1998-10-27
申请号：US242931
申请日：1994-05-16
CPC分类号： C12Q1/682 , C12Q1/6844 , C12Q1/6853 , C12Q1/6862 , C12Q1/689
摘要： A method is disclosed for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte. The method comprises (a) forming as a result of the presence of an analyte a single stranded polynucleotide comprising a target polynucleotide binding sequence flanked by first and second polynucleotide sequences that differ from the sequence of the analyte or a sequence complementary to the analyte sequence, (b) forming multiple copies of the single stranded polynucleotide, and (c) detecting the single stranded polynucleotide. Also disclosed is a method of producing at least one copy of a single stranded polynucleotide. The method comprises (a) forming in the presence of nucleoside triphosphates and template dependent polynucleotide polymerase an extension of a polynucleotide primer at least the 3'-end of which has at least a 10 base sequence hybridizable with a second sequence flanking the 3'-end of the single stranded polynucleotide, the second sequence being partially or fully complementary with at least a 10 base first sequence flanking the 5' end of the single stranded polynucleotide, (b) dissociating the extended polynucleotide primer and the single stranded polynucleotide, (c) repeating step a and (d) dissociating the extended polynucleotide primer and the copy of the single stranded polynucleotide.
摘要翻译： 公开了一种用于确定疑似含有分析物的样品中多核苷酸分析物的存在的方法。 该方法包括（a）作为分析物存在的结果形成单链多核苷酸，所述单链多核苷酸包含不同于分析物序列或与分析物序列互补的序列的第一和第二多核苷酸序列侧翼的靶多核苷酸结合序列， （b）形成多重拷贝的单链多核苷酸，和（c）检测单链多核苷酸。 还公开了生产单链多核苷酸的至少一个拷贝的方法。 该方法包括（a）在核苷三磷酸和模板依赖性多核苷酸聚合酶存在下形成多核苷酸引物的延伸，所述多核苷酸引物的至少3'末端具有至少10个碱基序列，可与3'- 单链多核苷酸的末端，第二个序列与单链多核苷酸的5'端侧翼的至少10个碱基第一序列部分或完全互补，（b）解离延伸的多核苷酸引物和单链多核苷酸，（c ）重复步骤a和（d）解离延伸的多核苷酸引物和单链多核苷酸的拷贝。
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