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1. 发明申请

US20110039720A1 DEVICE AND METHOD FOR PARALLEL QUANTITATIVE ANALYSIS OF MULTIPLE NUCLEIC ACIDS 审中-公开
标题翻译：用于多个核酸并行定量分析的装置和方法
公开(公告)号：US20110039720A1
公开(公告)日：2011-02-17
申请号：US12809412
申请日：2008-12-16
申请人： Erik R. Vossenaar , Derk J. W. Klunder , Hendrik R. Stapert , Henk Van Houten , Bob Van Gemen , Erik M. H. P. Van Dijk
发明人： Erik R. Vossenaar , Derk J. W. Klunder , Hendrik R. Stapert , Henk Van Houten , Bob Van Gemen , Erik M. H. P. Van Dijk
IPC分类号： C40B30/04 , C40B60/12
CPC分类号： C12Q1/6818 , C12Q1/6851 , C12Q2525/301 , C12Q2565/1015 , C12Q2565/549 , C12Q2561/113
摘要： The present invention relates to a process for conducting real-time PCR, and to a device for conducting the method of the present invention. The invention is especially suited for the simultaneous identification and quantification of nucleic acids present in a sample, e.g. a biological sample. Further, this invention describes a method for simultaneous quantitative analysis of multiple nucleic acid sequences in a single compartment by using an integrated nucleic acid microarray combined with a highly surface-specific readout device. The invention relates to a device wherein a surface which is either part of the chamber surface or a surface that is created in the reaction chamber, such as bead surface, is coated with capture probes and in the same chamber, a PCR reaction takes place.
摘要翻译：本发明涉及一种用于进行实时PCR的方法以及用于实施本发明的方法的装置。本发明特别适用于同时鉴定和定量样品中存在的核酸，例如，生物样品。此外，本发明描述了通过使用与高表面特异性读出装置组合的整合的核酸微阵列来同时定量分析单个隔室中的多个核酸序列的方法。本发明涉及一种装置，其中作为室表面的一部分的表面或在反应室中产生的表面（例如珠粒表面）的表面涂覆有捕获探针，并且在相同的室中进行PCR反应。

2. 发明授权

US5679553A Process for rendering a nucleic acid amplication reaction product incapable of being a target for further amplification, a diagnostic assay employing said process 失效
标题翻译：提供不能作为进一步扩增的靶标的核酸扩增反应产物的方法，采用所述方法的诊断测定法
公开(公告)号：US5679553A
公开(公告)日：1997-10-21
申请号：US519478
申请日：1995-08-25
申请人： Bob Van Gemen , Adriana Frederieke Schukkink
发明人： Bob Van Gemen , Adriana Frederieke Schukkink
IPC分类号： G01N33/50 , C07H21/00 , C07H21/04 , C12M1/00 , C12N15/09 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6865 , C12Q1/6848
摘要： A process for making the product of a nucleic acid amplification reaction, which amplification reaction employs one or more primer pairs, incapable of being a target for further amplification comprising: contacting the amplified product with an oligonucleotide capable of hybridizing to a stretch a nucleotides of the amplified product, said stretch being situated between the hybridisation sites of a pair of primers, under conditions which allow formation of a hybridisation complex between the oligonucleotide and the amplified product to occur, said oligonucleotide being modified in such a way that it.protects the double stranded part of the hybridisation complex hybridized to the oligonucleotide from degradation, and subjecting the hybridisation complex to a degradative treatment under circumstances such that at least the part of the hybridisation complex capable of hybridizing to the primers is degraded. An assay for diagnosing the presence of a nucleic acid sequence in a sample comprising carrying out the above-mentioned method and a kit for carrying out the above-mentioned method are also disclosed.
摘要翻译：一种制备核酸扩增反应产物的方法，该扩增反应采用一个或多个引物对，不能作为进一步扩增的靶，其包括：使扩增产物与能够与扩增产物，所述伸展位于一对引物的杂交位点之间，在允许在寡核苷酸和扩增产物之间形成杂交复合物的条件下发生，所述寡核苷酸以这样的方式进行修饰：保护双重杂交复合物的杂交部分与降解的寡核苷酸杂交，并且在使得能够与引物杂交的至少一部分杂交复合物降解的情况下使杂交复合物进行降解处理。还公开了用于诊断样品中存在核酸序列的测定法，包括进行上述方法和用于进行上述方法的试剂盒。

3. 发明授权

US06312928B1 Transcription based amplification of double stranded DNA targets 有权
公开(公告)号：US06312928B1
公开(公告)日：2001-11-06
申请号：US09554511
申请日：2000-05-16
申请人： Bob Van Gemen , Dianne Arnoldina Margaretha Van Strijp , Adriana Fredericke Schukkink
发明人： Bob Van Gemen , Dianne Arnoldina Margaretha Van Strijp , Adriana Fredericke Schukkink
IPC分类号： C12P1934
CPC分类号： C12Q1/6865 , C12Q2527/101
摘要： The present invention is directed to a transcription based amplification method for the amplification of DNA targets. With the method of the present invention an isothermal transcription based amplification method is provided for the amplification of double stranded DNA. The method of the present invention for the amplification of double stranded DNA comprises the steps of contacting said double stranded DNA with at least one oligonucleotide comprising a sequence complementary to a part of the first of the two DNA strands comprised in the double stranded DNA, said oligonucleotide further comprising the sequence of a promoter recognized by a RNA polymerase; a further olignucleotide comprising a sequence complementary to a part of the second strand comprised in the double stranded DNA; an enzyme having RNA dependent DNA polymerase activity; an enzyme having DNA dependent DNA polymerase activity; an enzyme having RNase H activity; an enzyme having RNA polymerase activity, and maintaining the thus created reaction mixture under the appropriate conditions for a sufficient amount of time for the amplification to take place. The method according to the invention is particularly useful for amplifying small DNA molecules, e.g. plasmid DNA. Surprisingly, transcription based amplification can start from double stranded DNA, without the need to treat the DNA with restriction enzymes or, what is more important, to separate the strands before hand by applying heat. All reagents conventionally used with isothermal transcription based amplification reactions can simply be used on starting material containing double stranded DNA, as if it where single stranded RNA. With the present invention it has now been found that essentially the same protocols can be used for isothermal transcription based amplification of double stranded DNA. The method is particularly useful for the detection of small DNA molecules of pathogenic microorganisms enabling diagnosis. In particular the circular HIV-1 DNA molecules that are formed during the replication of the HIV-1 virus can be detected by this method. Detection of such circular HIV-1 DNA molecules indicates active replication of the virus, which can be correlated with disease progression, i.e. development of AIDS.

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