公开(公告)号：EP1438437B1

公开(公告)日：2009-04-22

申请号：EP02802274.7

申请日：2002-10-25

申请人： Epigenomics AG

发明人： BERLIN, Kurt ,  BALLHAUSE, Matthias ,  GÜTIG, David

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q2565/518 ,  C12Q2531/113 ,  C12Q2523/125

摘要： A method for the detection of cytosine methylation in immobilised DNA samples is disclosed. In the first method step the genomic DNA is isolated from cells or other associated material and boned essentially irreversibly to a surface. The DNA bonded to the surface is then preferably treated with a bisulphite such that cytosine is converted into a base different from the base pairing relationship in the DNA duplex, whilst 5-methylcytosine remains unaltered. The used reagents are then removed in a washing step. Finally, selected sections of the immobilised DNA are amplified in a polymerase reaction and the amplificate examined with regard to the sequence thereof.

公开(公告)号：EP1438437A2

公开(公告)日：2004-07-21

申请号：EP02802274.7

申请日：2002-10-25

申请人： Epigenomics AG

发明人： BERLIN, Kurt ,  BALLHAUSE, Matthias ,  GÜTIG, David

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q2565/518 ,  C12Q2531/113 ,  C12Q2523/125

摘要： A method for the detection of cytosine methylation in immobilised DNA samples is disclosed. In the first method step the genomic DNA is isolated from cells or other associated material and boned essentially irreversibly to a surface. The DNA bonded to the surface is then preferably treated with a bisulphite such that cytosine is converted into a base different from the base pairing relationship in the DNA duplex, whilst 5-methylcytosine remains unaltered. The used reagents are then removed in a washing step. Finally, selected sections of the immobilised DNA are amplified in a polymerase reaction and the amplificate examined with regard to the sequence thereof.

公开(公告)号：EP1412524A2

公开(公告)日：2004-04-28

申请号：EP02747242.2

申请日：2002-06-27

申请人： Epigenomics AG

发明人： GÜTIG, David

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6869 ,  C12Q1/6858 ,  C12Q2563/107 ,  C12Q2565/101 ,  C12Q2531/113 ,  C12Q2523/125

摘要： The invention relates to a method for detecting specific dinucleotides in a DNA-sample. A polymerase-chain reaction (PCR) is carried out by using a) a nucleotide, which is part of the dinucleotide which is to be detected, wherein an adequate quantity thereof is marked by a donor-fluorphore and b) another nucleotide, which is part of the dinucleotide which is to be detected, wherein an adequate quantity thereof is marked with an acceptor-fluorophore. Said method determines or quantifies the presence of the dinucleotide by measuring the dimensions of the fluorescence resonance energy transfer (FRET) between the donor- and acceptor-fluorophore.

公开(公告)号：EP1644521A1

公开(公告)日：2006-04-12

申请号：EP04740437.1

申请日：2004-06-29

申请人： Epigenomics AG

发明人： GÜTIG, David

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q2521/331

摘要： The present invention concerns a method for investigating cytosine methylations in DNA sequences. By this means, the DNA to be investigated is reacted with a cytidine deaminase which deaminates cytidine more rapidly than 5-methylcytidine. Cytosine is converted to uracil by the conversion, whereas 5-methylcytosine remains essentially unchanged. The enzymatically pretreated DNA is preferably amplified and can be analyzed by different methods. The method according to the invention is particularly suitable for the diagnosis of cancer diseases and other disorders associated with a change in the methylation status, as well as for the prognosis of undesired drug effects.

公开(公告)号：EP1521635A1

公开(公告)日：2005-04-13

申请号：EP03760576.3

申请日：2003-06-23

申请人： Epigenomics AG

发明人： GÜTIG, David

IPC分类号： B01L3/00 ,  G01N1/00

CPC分类号： G01N35/1097 ,  B01L3/502 ,  B01L3/5027 ,  B01L2300/0816 ,  B01L2300/0861 ,  B01L2400/0478 ,  B01L2400/0622 ,  B01L2400/0644

摘要： The invention relates to a device for handling liquids contained in at least two channels. Said device consists of one component, referred to subsequently as a channel support, in which the channels extend in a stellate manner from a cone and a second component, subsequently referred to as a pump element, which contains at least one chamber with a variable volume and has a cone-shaped end with at least one integrated lateral opening that is directed towards at least one of the volumes. The cones of the components are fitted into one another in such a way as to permit contact between at least one opening of the pump element and at least one of the channels in the channel support, by rotating the components in relation to one another about the cone axes, the formation of a connection between at least one volume and at least one channel and the closure of other openings and channels by cone surfaces.

公开(公告)号：EP1423528B1

公开(公告)日：2008-01-02

申请号：EP01980315.4

申请日：2001-09-01

申请人： Epigenomics AG

发明人： OLEK, Alexander ,  PIEPENBROCK, Christian ,  BERLIN, Kurt ,  GÜTIG, David

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q1/6883 ,  C12Q2600/154

摘要： The invention relates to a method for detecting the degree of methylation of a defined cytosine in the sequence context 5′-CpG-3′ of a genomic DNA sample. The first stage involves chemically treating the genomic DNA in such a way that the cytosine bases, but not the 5-methylcytosine bases, are converted into uracil. Parts of the genomic DNA containing the defined cytosine are then amplified. The amplified parts are given a detectable mark and the extent of the hybridisation of the amplified parts on the two classes of oligonucleotides is then determined by detecting the mark of the amplified parts. The degree of methylation of the defined cytosine in the genomic DNA sample can be deduced on the basis of the relationship between the marks detected on the two classes of oligonucleotides following the hybridisation.

公开(公告)号：EP1733054A2

公开(公告)日：2006-12-20

申请号：EP05737974.5

申请日：2005-04-06

申请人： Epigenomics AG

发明人： GÜTIG, David ,  HABIGHORST, Dirk ,  KLUTH, Antje ,  SCHMITT, Armin ,  SCHUSTER, Matthias ,  SCHWOPE, Ina

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6823 ,  C12Q2565/101 ,  C12Q2535/131 ,  C12Q2545/114 ,  C12Q2523/125

摘要： The method according to the invention concerns a method for the quantification of two different variations of a DNA sequence. Particularly, the invention relates to a quantification of methylated DNA. For this purpose, the DNA to be investigated is first converted so that cytosine is converted to uracil, while 5-methylcytosine remains unchanged. Then the converted DNA is amplified by means of a real-time PCR. However, probes are utilized, one of which is specific for the methylated status and one for the unmethylated status of the DNA. The degree of methylation of the investigated DNA can be calculated from the ratio of the signal intensities of the probes or from the Ct values. The method according to the invention is particularly suitable for the diagnosis and prognosis of cancer and other disorders associated with a change in the methylation status, as well as for the prediction of drug interactions.

公开(公告)号：EP1423528A2

公开(公告)日：2004-06-02

申请号：EP01980315.4

申请日：2001-09-01

申请人： Epigenomics AG

发明人： OLEK, Alexander ,  PIEPENBROCK, Christian ,  BERLIN, Kurt ,  GÜTIG, David

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q1/6883 ,  C12Q2600/154

摘要： The invention relates to a method for detecting the degree of methylation of a defined cytosine in the sequence context 5′-CpG-3′ of a genomic DNA sample. The first stage involves chemically treating the genomic DNA in such a way that the cytosine bases, but not the 5-methylcytosine bases, are converted into uracil. Parts of the genomic DNA containing the defined cytosine are then amplified. The amplified parts are given a detectable mark and the extent of the hybridisation of the amplified parts on the two classes of oligonucleotides is then determined by detecting the mark of the amplified parts. The degree of methylation of the defined cytosine in the genomic DNA sample can be deduced on the basis of the relationship between the marks detected on the two classes of oligonucleotides following the hybridisation.

摘要翻译： 确定存在于DNA（I）的基因组样品中的基序5'-CpG-3'中的特定胞嘧啶的甲基化程度是新的。 确定存在于DNA（I）的基因组样品中的基序5'-CpG-3'中的特定胞嘧啶的甲基化程度是新的。 将样品化学处理以将胞嘧啶（C）转化为尿嘧啶，然后将含有目标C的（I）部分扩增以形成标记的扩增子（II）。 （II）与寡核苷酸和/或肽 - 核酸（PNA）寡聚体（A）的两个类别分别与至少一个成员杂交，并且从（II）上的标记确定与两个等级的杂交程度。 从与（A）两类杂交的标记的比例，计算甲基化程度。 还包括以下独立权利要求：（1）包含亚硫酸氢盐，用于扩增的引物寡核苷酸和/或（A））的试剂盒，优选固定在固相上，加上说明书; （2）在说明书中未给出的序列（1） - （40712）的化学预处理的DNA（B）至少包含18个碱基的核酸; （3）用于新方法的寡核苷酸或PNA寡聚体（OG），其包含能够与（B）杂交的至少9个核苷酸（nt）的序列; （4）OG组; （5）至少两个用于扩增至少一个（B）或其片段的引物寡核苷酸的组; （6）用于确定（B）中的C甲基化和/或单核苷酸多态性程度的寡核苷酸探针组，其包含至少10个OG; （7）通过将OG连接到固相，制备不同OG的载体结合阵列，用于分析与（B）的甲基化状态相关的疾病; 和（8）由（7）制成的阵列。