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1. 发明公开

EP2765202A1 Methods of nonspecific target capture of nucleic acids 有权
标题翻译：维尔法赫尔·祖尔·尼希茨菲西岑Zielerfassung vonNukleinsäuren
公开(公告)号：EP2765202A1
公开(公告)日：2014-08-13
申请号：EP13160839.0
申请日：2007-08-01
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Majlessi, Mehrdad R.
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12N15/1006 , C12N15/11 , C12N2310/18 , C12N2310/31 , C12N2310/321 , C12N2310/351 , C12N2330/30 , C12Q1/6806 , C12Q1/6834 , C12Q1/70 , C12Q2565/519 , C12Q2565/518
摘要： Methods for capturing a target nucleic acid from a sample by using a capture probe that binds nonspecifically to the target nucleic acid and binds specifically to an immobilized probe via a specific binding pair that has one member on the capture probe and one member on the immobilized probe are disclosed. Compositions that include a capture probe that binds nonspecifically to a target nucleic acid and specifically to an immobilized probe via binding of members of a specific binding pair in a solution phase of a reaction mixture are disclosed.
摘要翻译：通过使用捕获探针非特异性结合靶核酸并通过在捕获探针上具有一个成员的特异性结合对与固定化探针上的一个成员特异性结合固定的探针的方式从样品中捕获靶核酸的方法被披露。公开了包含捕获探针的组合物，其通过结合反应混合物的溶液相中的特异性结合对的成员非特异性地结合靶核酸，特异性结合固定的探针。

2. 发明公开

EP2345740A1 Tagged oligonucleotides and their use in nucleic acid amplification methods 有权
标题翻译：在Nukleinsäureverstärkungsverfahren的Markierte Oligonukleotide und ihre Verwendung
公开(公告)号：EP2345740A1
公开(公告)日：2011-07-20
申请号：EP10015034.1
申请日：2007-06-06
申请人： Gen-Probe Incorporated
发明人： Becker, Michael M. , Livezey, Kristin W. , Lam, Wai-Chung
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6853 , C12Q1/6848 , C12Q1/6865 , C12Q2525/301 , C12Q2525/186 , C12Q2525/155 , C12Q2525/161
摘要： A pre-amplification reaction mixture for selective amplification of one or more target nucleic acid sequences, said reaction mixture comprising: a tagged oligonucleotide comprising first and second regions, said first region comprising a target hybridizing sequence hybridized to a target region contained at a 3'-end of one or more target nucleic acid sequences present in said reaction mixture and said second region comprising a tag sequence situated 5' to said target hybridizing sequence; a first oligonucleotide comprising a hybridizing sequence which hybridizes to a 3'-end of the complement of one or more of said target nucleic acid sequences; and a second oligonucleotide comprising a hybridizing sequence which hybridizes to the complement of said tag sequence, wherein said reaction mixture is substantially free of an active form of said tagged oligonucleotide which is not hybridized to said target region contained in said one or more target nucleic acid sequences present in said reaction mixture, wherein said active form of said tagged oligonucleotide has an available target hybridizing sequence for hybridization to said target region present in a non-target nucleic acid added to said reaction mixture, and wherein said reaction mixture does not comprise a nucleic acid polymerase capable of extending any of said oligonucleotides in a template-dependent manner.
摘要翻译：用于选择性扩增一个或多个靶核酸序列的预扩增反应混合物，所述反应混合物包含：包含第一和第二区域的标记寡核苷酸，所述第一区域包含与包含在3'端的靶区域杂交的靶杂交序列，存在于所述反应混合物中的一个或多个靶核酸序列，并且所述第二区包含位于所述靶杂交序列5'的标签序列; 第一寡核苷酸，其包含与一个或多个所述靶核酸序列的补体的3'-末端杂交的杂交序列; 以及第二寡核苷酸，其包含与所述标签序列的互补序列杂交的杂交序列，其中所述反应混合物基本上不含所述标记的寡核苷酸的活性形式，其与所述一种或多种靶核酸中包含的所述靶标区域不杂交存在于所述反应混合物中的序列，其中所述标记的寡核苷酸的所述活性形式具有用于与存在于添加到所述反应混合物中的非靶核酸中的所述靶区杂交的可用靶杂交序列，并且其中所述反应混合物不包含能够以模板依赖的方式扩展任何所述寡核苷酸的核酸聚合酶。

3. 发明公开

EP2071031A1 Single-primer nucleic acid amplification methods 失效
标题翻译： EINFACH-Primernukleinsäuren-Erweiterungsverfahren
公开(公告)号：EP2071031A1
公开(公告)日：2009-06-17
申请号：EP08005114.7
申请日：2005-08-26
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Lam, Wai-Chung , Livezey, Kristin W. , Brentano, Steven T. , Kolk, Daniel P. , Schroder, Astrid R. W.
IPC分类号： C12P19/34 , C07H21/04
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2537/163 , C12Q2521/325 , C12Q2521/107 , C12Q2525/186 , C12Q2533/101 , C12Q2525/143
摘要： The present invention relates, in one aspect, to a method of synthesizing multiple copies of a target sequence, said method comprising the steps of: (A) treating a target nucleic acid comprising an RNA target sequence with: (1) a priming oligonucleotide which hybridizes to the 3'-end of said target sequence such that a primer extension reaction can be initiated therefrortt, wherein said priming oligonucleotide has a cap hybridized to a 3'-end thereof prior to hybridizing to said target sequence, said cap comprising a base region which is complementary to at least 3 nucleotides at the 3'-end of said priming oligonucleotide, wherein the 5'-terminal base of said cap is complementary to the 3'-terminal base of said priming oligonucleotide, and wherein said cap is modified to prevent the initiation of DNA synthesis therefrom; and (2) a binding molecule which binds to said target nucleic acid adjacent to or near the 5'-end of said target sequence; (B) extending said priming oligonucleotide in a primer extension reaction with a DNA polymerase to give a DNA primer extension product complementary to said target sequence, said DNA primer extension product having a 3'-end which is determined by said binding molecule and which is complementary to the 5'-end of said target sequence; (C) separating said DNA primer extension product from said target sequence using an enzyme which selectively degrades said target sequence; (D) treating said DNA primer extension product with a promoter oligonucleotide comprising first and second regions, said first region hybridizing to a 3'-region of said DNA primer extension product to form a promoter oligonucleotide:DNA primer extension product hybrid, and said second region being a promoter for an RNA polymerase and situated 5' to said first region, wherein said promoter oligonucleotide is modified to prevent the initiation of DNA synthesis therefrom; (E) extending the 3'-end of said DNA primer extension product in said promoter oligonucleotide:DNA primer extension product hybrid to add a sequence complementary to said second region of said promoter oligonucleotide; and (F)
transcribing from said promoter oligonucleotide:DNA primer extension product hybrid multiple RNA products complexnentary to said DNA primer extension product using an RNA polymerase which recognizes said promoter and initiates, transcription therefrom wherein the base sequences of said RNA products are substantially identical to the base sequence of said target sequence.
摘要翻译：本发明一方面涉及一种合成靶序列多拷贝的方法，所述方法包括以下步骤：（A）用下列步骤处理包含RNA靶序列的靶核酸：（1）引物寡核苷酸与所述靶序列的3'末端杂交，使得可引发引物延伸反应，其中所述引物寡核苷酸在与所述靶序列杂交之前具有与其3'-末端杂交的帽，所述帽包含碱其与所述引发寡核苷酸的3'末端至少3个核苷酸互补，其中所述帽的5'末端碱基与所述引物寡核苷酸的3'末端碱基互补，并且其中所述帽被修饰以防止从其中开始DNA合成; 和（2）结合分子，所述结合分子与所述靶序列的5'末端相邻或相近的所述靶核酸结合; （B）在与DNA聚合酶的引物延伸反应中延伸所述引物寡核苷酸，得到与所述靶序列互补的DNA引物延伸产物，所述DNA引物延伸产物具有由所述结合分子确定的3'末端，与所述靶序列的5'末端互补; （C）使用选择性降解所述靶序列的酶分离所述DNA引物延伸产物与所述靶序列; （D）用包含第一和第二区域的启动子寡核苷酸处理所述DNA引物延伸产物，所述第一区域与所述DNA引物延伸产物的3'区域杂交以形成启动子寡核苷酸：DNA引物延伸产物杂交体，所述第二区域区域是RNA聚合酶的启动子，并且位于所述第一区域5'处，其中所述启动子寡核苷酸被修饰以防止从其中开始DNA合成; （E）在所述启动子寡核苷酸中扩展所述DNA引物延伸产物的3'-末端：DNA引物延伸产物杂交物，以加入与所述启动子寡核苷酸的所述第二区域互补的序列; 和（F）从所述启动子寡核苷酸转录：使用识别所述启动子并引发转录的RNA聚合酶与所述DNA引物延伸产物复合的DNA引物延伸产物杂合多RNA产物，其中所述RNA产物的碱基序列基本上等同于所述靶序列的碱基序列。

4. 发明公开

EP2017356A3 Tagged oliggonucleotides and their use in nucleic acid amplification methods 失效
标题翻译：标记的寡核苷酸及其在核酸扩增方法中的用途
公开(公告)号：EP2017356A3
公开(公告)日：2009-05-06
申请号：EP08005731.8
申请日：2008-06-12
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Livezy, Kristin W. , Lam, Wai-Chung
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6853 , C12Q1/6848 , C12Q1/6865 , C12Q2525/301 , C12Q2525/186 , C12Q2525/155 , C12Q2525/161
摘要： The present invention relates to a method for the selective amplification of at least one target nucleic acid sequence from a nucleic acid sample, said method comprising the steps of (a) treating a nucleic acid sample comprising a target nucleic acid sequence with a tagged oligonucleotide comprising first and second regions, said first region comprising a target hybridizing sequence which hybridizes to a 3'-end of said target nucleic acid sequence and said second region comprising a tag sequence situated 5' to said target hybridizing sequence, wherein said second region does not stably hybridize to a target nucleic acid containing said target nucleic acid sequence; (b)reducing in said nucleic acid sample the effective concentration of unhybridized tagged oligonucleotide having an active form in which a target hybridizing sequence of said unhybridized tagged oligonucleotide is available for hybridization to said target nucleic acid sequence; and (c) producing amplification products in a nucleic acid amplification reaction using first and second oligonucleotides, wherein said first oligonucleotide comprises a hybridizing sequence which hybridizes to a 3'-end of the complement of said target nucleic acid sequence and said second oligonucleotide comprises a hybridizing sequence which hybridizes to the complement of said tag sequence, wherein said second oligonucleotide does stably hybridize to a target nucleic acid containing said target nucleic acid sequence, wherein each of said amplification products comprises a base sequence which is substantially identical or complementary to the base sequence of said target nucleic acid sequence and further comprises a base sequence which is substantially identical or complementary to all or a portion of said tag sequence, and wherein step (b) comprises inactivating unhybridized tagged oligonucleotide so that said unhybridized tagged oligonucleotide does not stably hybridize to said target nucleic acid sequence during step (c).
摘要翻译：本发明涉及用于从核酸样品中选择性扩增至少一种靶核酸序列的方法，所述方法包括以下步骤：（a）用标记的寡核苷酸处理包含靶核酸序列的核酸样品，所述标记的寡核苷酸包含第一和第二区域，所述第一区域包含与所述靶核酸序列的3'-末端杂交的靶杂交序列，所述第二区域包含位于所述靶杂交序列5'的标签序列，其中所述第二区域不包含与含有所述靶核酸序列的靶核酸稳定地杂交; （b）在所述核酸样品中还原具有活性形式的未杂交标记寡核苷酸的有效浓度，其中所述未杂交标记寡核苷酸的靶杂交序列可用于与所述靶核酸序列杂交; 和（c）在使用第一和第二寡核苷酸的核酸扩增反应中产生扩增产物，其中所述第一寡核苷酸包含与所述靶核酸序列的互补序列的3'-末端杂交的杂交序列，并且所述第二寡核苷酸包含杂交序列，其与所述标签序列的互补序列杂交，其中所述第二寡核苷酸稳定地与含有所述靶核酸序列的靶核酸杂交，其中每种所述扩增产物包含与所述碱基本相同或互补的碱基序列所述靶标核酸序列还包含与所述标签序列的全部或部分基本相同或互补的碱基序列，并且其中步骤（b）包括使未杂交的标记的寡核苷酸失活，使得所述未杂交的标记的寡核苷酸不稳定杂交到所述焦油在步骤（c）期间获得核酸序列。

5. 发明公开

EP2017356A2 Tagged oliggonucleotides and their use in nucleic acid amplification methods 失效
标题翻译：在Nukleinsäureverstärkungsverfahren的Markierte Oligonukleotide und ihre Verwendung
公开(公告)号：EP2017356A2
公开(公告)日：2009-01-21
申请号：EP08005731.8
申请日：2008-06-12
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Livezy, Kristin W.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6853 , C12Q1/6848 , C12Q1/6865 , C12Q2525/301 , C12Q2525/186 , C12Q2525/155 , C12Q2525/161
摘要： The present invention relates to a method for the selective amplification of at least one target nucleic acid sequence from a nucleic acid sample, said method comprising the steps of (a) treating a nucleic acid sample comprising a target nucleic acid sequence with a tagged oligonucleotide comprising first and second regions, said first region comprising a target hybridizing sequence which hybridizes to a 3'-end of said target nucleic acid sequence and said second region comprising a tag sequence situated 5' to said target hybridizing sequence, wherein said second region does not stably hybridize to a target nucleic acid containing said target nucleic acid sequence; (b)reducing in said nucleic acid sample the effective concentration of unhybridized tagged oligonucleotide having an active form in which a target hybridizing sequence of said unhybridized tagged oligonucleotide is available for hybridization to said target nucleic acid sequence; and (c) producing amplification products in a nucleic acid amplification reaction using first and second oligonucleotides, wherein said first oligonucleotide comprises a hybridizing sequence which hybridizes to a 3'-end of the complement of said target nucleic acid sequence and said second oligonucleotide comprises a hybridizing sequence which hybridizes to the complement of said tag sequence, wherein said second oligonucleotide does stably hybridize to a target nucleic acid containing said target nucleic acid sequence, wherein each of said amplification products comprises a base sequence which is substantially identical or complementary to the base sequence of said target nucleic acid sequence and further comprises a base sequence which is substantially identical or complementary to all or a portion of said tag sequence, and wherein step (b) comprises inactivating unhybridized tagged oligonucleotide so that said unhybridized tagged oligonucleotide does not stably hybridize to said target nucleic acid sequence during step (c).
摘要翻译：本发明涉及从核酸样品中选择性扩增至少一种靶核酸序列的方法，所述方法包括以下步骤：（a）用包含目标核酸序列的核酸样品处理含有标记的寡核苷酸的方法，所述标记的寡核苷酸包含第一和第二区域，所述第一区域包含与所述靶核酸序列的3'-末端杂交的靶杂交序列，所述第二区域包含位于所述靶杂交序列5'的标签序列，其中所述第二区域不与含有所述靶核酸序列的靶核酸稳定杂交; （b）在所述核酸样品中减少具有活性形式的未杂交标记的寡核苷酸的有效浓度，其中所述未杂交标记的寡核苷酸的靶杂交序列可用于与所述靶核酸序列杂交; 和（c）使用第一和第二寡核苷酸在核酸扩增反应中产生扩增产物，其中所述第一寡核苷酸包含与所述靶核酸序列的互补序列的3'-末端杂交的杂交序列，所述第二寡核苷酸包含与所述标签序列的互补序列杂交的杂交序列，其中所述第二寡核苷酸与含有所述靶核酸序列的靶核酸稳定杂交，其中每个所述扩增产物包含基本上与碱基相同或互补的碱基序列所述靶核酸序列的序列还包含与所述或一部分所述标签序列基本相同或互补的碱基序列，并且其中步骤（b）包括使未杂交的标记的寡核苷酸失活，使得所述未杂交的标记的寡核苷酸不稳定杂交到焦油在步骤（c）期间获得核酸序列。

6. 发明公开

EP0851033A1 Target-triggered amplification 失效
标题翻译： ZielsequenzabhängigeAmplifizierung
公开(公告)号：EP0851033A1
公开(公告)日：1998-07-01
申请号：EP97310550.5
申请日：1997-12-23
申请人： Gen-Probe Incorporated
发明人： Stull, Paul D. , Myers, Kristi K. , Becker, Michael M.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/682 , C12Q1/6865 , C12Q2525/301 , C12Q2525/161 , C12Q2525/143
摘要： The present invention features "promoter-sequestered" oligonucleosides and the use of such oligonucleosides to achieve "target-triggered" amplification. A promoter-sequestered oligonucleoside contains a contiguous nucleic acid sequence which forms a stem-loop structure in the absence of a target sequence. The stem-loop structure contains a single-stranded loop region and a double-stranded stem region. The single-stranded loop contains all, or a portion of, an RNA polymerase promoter sequence. The stem is produced from two substantially complementary nucleic acid sequences able to form an intramolecular hybrid. The secondary structure of the stem decreases the accessibility of the loop promoter sequence to form a functional double-stranded promoter.
摘要翻译：本发明的特征在于“启动子隔离”寡核苷，并且使用这些寡核苷来实现“目标触发”扩增。启动子螯合的寡核苷酸含有连续的核酸序列，其在不存在靶序列的情况下形成茎 - 环结构。茎环结构包含单链环区和双链茎区。单链环包含RNA聚合酶启动子序列的全部或部分。茎由两个能够形成分子内杂交体的基本互补的核酸序列产生。茎的二级结构降低环型启动子序列的可及性以形成功能性双链启动子。

7. 发明授权

EP2765202B1 Methods of nonspecific target capture of nucleic acids 有权
标题翻译：一种用于核酸的非特异性靶检测方法
公开(公告)号：EP2765202B1
公开(公告)日：2017-02-01
申请号：EP13160839.0
申请日：2007-08-01
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Majlessi, Mehrdad R.
IPC分类号： C12Q1/68
CPC分类号： C12N15/1006 , C12N15/11 , C12N2310/18 , C12N2310/31 , C12N2310/321 , C12N2310/351 , C12N2330/30 , C12Q1/6806 , C12Q1/6834 , C12Q1/70 , C12Q2565/519 , C12Q2565/518

8. 发明授权

EP1970454B1 Single-primer nucleic acid amplification methods 失效
标题翻译：只要核酸引物扩大进程
公开(公告)号：EP1970454B1
公开(公告)日：2012-09-19
申请号：EP08004311.0
申请日：2005-08-26
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Lam, Wai-Chung , Livezey, Kristin W. , Brentano, Steven T. , Kolk, Daniel P. , Schroder, Astrid R.W.
IPC分类号： C12P19/34 , C07H21/04 , C12Q1/68
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2537/163 , C12Q2521/325 , C12Q2521/107 , C12Q2525/186 , C12Q2533/101 , C12Q2525/143

9. 发明授权

EP1975242B1 Single-primer nucleic acid amplification methods 失效
标题翻译：只要核酸引物扩大进程
公开(公告)号：EP1975242B1
公开(公告)日：2011-03-02
申请号：EP08005147.7
申请日：2005-08-26
申请人： GEN-PROBE INCORPORATED
发明人： Becker, Michael M. , Lam, Wai-Chung , Livezey, Kristin W. , Brentano, Steven T. , Kolk, Daniel P. , Schroder, Astrid R.W.
IPC分类号： C12Q1/68
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2537/163 , C12Q2521/325 , C12Q2521/107 , C12Q2525/186 , C12Q2533/101 , C12Q2525/143

10. 发明公开

EP2243832A3 Sample preparation method incorporating an alkaline shock 审中-公开
标题翻译：样品制备方法用碱休克
公开(公告)号：EP2243832A3
公开(公告)日：2011-01-05
申请号：EP10006190.2
申请日：2006-02-17
申请人： Gen-Probe Incorporated
发明人： Gao, Kui , Becker, Michael M. , Wu, Wen , Linnen, Jeffrey M.
IPC分类号： C12N15/10
CPC分类号： C12N15/1006 , C12N15/1013 , C12Q1/6806 , C12Q1/689 , C12Q2527/119
摘要： Method of preparing a biological sample appropriate for use in a subsequent in vitro nucleic acid amplification reaction. The method involves a rapid, transient exposure to alkaline conditions which can be achieved by mixing an alkaline solution with a pH-buffered solution that includes a detergent and the biological sample to be tested for the presence of particular nucleic acid species using in vitro amplification. The invention method advantageously can improve detection of some target nucleic acids without substantially compromising detectability of others. The method is particularly useful for simultaneously preparing RNA and DNA templates that can be used in multiplex amplification reactions.

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