公开(公告)号：WO2009086218A3

公开(公告)日：2009-09-03

申请号：PCT/US2008087859

申请日：2008-12-19

Applicant: GEN PROBE INC ,  BECKER MICHAEL M ,  GAO KUI ,  LAM WAI-CHUNG

Inventor： BECKER MICHAEL M ,  GAO KUI ,  LAM WAI-CHUNG

IPC: C12Q1/68

CPC classification number: C12Q1/689 ,  C12Q2600/16 ,  C12Q2545/114 ,  C12Q2545/101 ,  C12Q2531/113

Abstract: Method of detecting methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) in a nucleic acid coamplification assay. The invention advantageously reduces the incidence of false-positive MRSA determinations in real-time assays by requiring satisfaction of a threshold criterion that excludes certain co-infections from the MRSA determination. The invention further provides for determination of MSSA, even when the MSSA is present in combination with methicillin-resistant coagulase-negative (MR-CoNS) bacteria at high or low levels.

Abstract translation: 在核酸共扩增测定中检测耐甲氧西林金黄色葡萄球菌（MRSA）和甲氧西林敏感性金黄色葡萄球菌（MSSA）的方法。 本发明有利地通过要求满足从MRSA测定中排除某些共感染的阈值标准来降低实时分析中假阳性MRSA测定的发生率。 本发明还提供MSSA的测定，即使MSSA与高水平或低水平的耐甲氧西林凝固酶阴性（MR-CoNS）细菌组合存在。

公开(公告)号：WO2008108843A3

公开(公告)日：2008-10-30

申请号：PCT/US2007063103

申请日：2007-03-01

Applicant: GEN PROBE INC ,  BECKER MICHAEL M ,  LAM WAI-CHUNG ,  LIVEZEY KRISTIN W ,  BRENTANO STEVEN T ,  KOLK DANIEL P ,  SCHRODER ASTRID R W

Inventor： BECKER MICHAEL M ,  LAM WAI-CHUNG ,  LIVEZEY KRISTIN W ,  BRENTANO STEVEN T ,  KOLK DANIEL P ,  SCHRODER ASTRID R W

IPC: C12Q1/68 ,  C07H21/02

CPC classification number: C12Q1/6865 ,  C12Q2533/101 ,  C12Q2525/186 ,  C12Q2521/107

Abstract: Novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic are disclosed (/. e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, methods of nucleic acid amplification are disclosed which are robust and efficient, while reducing the appearance of side-products. In general, the methods use priming oligonucleotides that target only one sense of a target nucleic acid, a promoter oligonucleotide modified to prevent polymerase extension from its 3 '-terminus and, optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or substantially eliminating the formation of side-products. The disclosed methods minimizes or substantially eliminate the emergence of side-products, thus providing ahigh level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The disclosed methods minimize or substantially eliminate this problem, thus providing enhanced levels of sensitivity.

Abstract translation: 公开了合成自动催化的靶核酸序列的多个拷贝的新方法（例如，能够自动循环而不需要修饰反应条件如温度，pH或离子强度，并且使用一个循环的产物 在下一个）。 特别地，公开了鲁棒和有效的核酸扩增方法，同时减少副产物的外观。 通常，所述方法使用仅靶向靶核酸的引物寡核苷酸，修饰以阻止聚合酶从其3'末端延伸的启动子寡核苷酸，以及任选的终止引物延伸反应的手段以扩增RNA或 DNA分子在体外，同时减少或基本上消除副产物的形成。 所公开的方法最小化或基本上消除副产物的出现，从而提供高水平的特异性。 此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。 所公开的方法最小化或基本上消除了该问题，从而提供了增强的灵敏度水平。

公开(公告)号：WO2006026388A3

公开(公告)日：2007-01-18

申请号：PCT/US2005030329

申请日：2005-08-26

Applicant: GEN PROBE INC ,  BECKER MICHAEL M ,  LAM WAI-CHUNG ,  LIVEZEY KRISTIN W ,  BRENTANO STEVEN T ,  KOLK DANIEL P ,  SCHRODER ASTRID R W

Inventor： BECKER MICHAEL M ,  LAM WAI-CHUNG ,  LIVEZEY KRISTIN W ,  BRENTANO STEVEN T ,  KOLK DANIEL P ,  SCHRODER ASTRID R W

IPC: C12P19/34 ,  C07H21/04

CPC classification number: C12P19/34 ,  C12Q1/6865 ,  C12Q2537/163 ,  C12Q2521/325 ,  C12Q2521/107 ,  C12Q2525/186 ,  C12Q2533/101 ,  C12Q2525/143

Abstract: The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side-products. The method uses only one primer, the "priming oligonucleotide," a 3'blocked promoter oligonucleotide and optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or eliminating the formation of side-products. The method of the present invention minimizes or eliminates the emergence of side-products, thus providing a high level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or eliminates this problem, thus providing an enhanced level of sensitivity.

Abstract translation: 本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种 循环在下一个）。 特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少副产物的外观。 该方法仅使用一种引物“引物寡核苷酸”，3'封闭的启动子寡核苷酸和任选的终止引物延伸反应的手段，以体外扩增RNA或DNA分子，同时减少或消除副产物的形成 。 本发明的方法使副产物的出现最小化或消除，从而提供高水平的特异性。 此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。 本发明使这个问题最小化或消除了这一点，从而提供了增强的灵敏度。