摘要:
The present invention provides a method for determination of presence or absence of an epithelial cancer-derived cell in a biological sample obtained from a subject comprising the steps of: extracting DNA from the biological sample, analyzing methylation status of a CpG site located in at least one region represented by base sequences SEQ ID NOs: 1, 2, 3 and 4 in the DNA obtained from the step of extracting, and determining presence or absence of the epithelial cancer-derived cell in the biological sample based on an analysis result obtained from the step of analyzing.
摘要翻译:本发明提供一种用于确定从受试者获得的生物样品中存在或不存在上皮癌衍生细胞的方法,包括以下步骤:从生物样品中提取DNA,分析至少位于至少一个CpG位点的甲基化状态 由从提取步骤获得的DNA中的碱基序列SEQ ID NO:1,2,3和4表示的一个区域,并且基于从生物样品中获得的分析结果确定生物样品中上皮癌来源的细胞的存在或不存在 分析的步骤。
摘要:
Success or failure of unmethylated cytosine conversion treatment is determined by using a first primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence not containing cytosine in the nucleotide sequence of biological DNA that is subject to an unmethylated cytosine conversion treatment and a second primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence in which cytosine in a nucleotide sequence containing cytosine and not containing a CpG site is converted into a base other than cytosine, in the nucleotide sequence of biological DNA.
摘要:
Success or failure of unmethylated cytosine conversion treatment is determined by using a first primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence not containing cytosine in the nucleotide sequence of biological DNA that is subject to an unmethylated cytosine conversion treatment and a second primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence in which cytosine in a nucleotide sequence containing cytosine and not containing a CpG site is converted into a base other than cytosine, in the nucleotide sequence of biological DNA.
摘要:
The present invention provides a sample treatment solution for preparing a sample for DNA methylation which can achieve stable detection results in detection of DNA methylation and be easily pretreated, comprising an aqueous solution containing a protease.
摘要:
The present invention provides a method of analyzing methylated DNA, comprising steps of: (A) treating a DNA-containing sample with a restriction enzyme to obtain a sample containing a DNA fragment; (B) concentrating methylated DNA contained in the sample obtained in step (A) to obtain a methylated DNA concentrate; (C) subjecting the methylated DNA concentrate obtained in step (B) and a primer set to nucleic acid amplification reaction, wherein the primer set performs the nucleic acid amplification reaction in step (C) by using a template DNA which does not have a CpG site; (D) detecting an amplification product obtained in step (C); (E) judging whether the methylated DNA concentrate obtained in step (B) is appropriate as a sample for detection of methylated DNA, on the basis of the detection result of the amplification product in step (D); and (F) analyzing the methylated DNA contained in the methylated DNA concentrate.
摘要:
The present invention relates to a hybridoma producing an anti-methylated DNA antibody, obtained by cell fusion of an antibody-producing cell obtained from an animal immunized with an antigen containing 5′-(5-methyl-2′-deoxycytidine-3′-phospho)-2′-deoxyguanosine 3′-phosphate with a myeloma cell. The present invention also relates to a monoclonal antibody produced by the hybridoma and a method for immunoprecipitation of a methylated DNA using the antibody.
摘要:
The method for detecting a methylated cytosine of the present invention comprises the steps of: hybridizing a sample DNA with an oligonucleotide which can hybridize with a region of the sample DNA containing a cytosine suspected of being methylated and has an a basic site at the position complementary to the cytosine; reacting the hybridized sample DNA obtained in the previous step with an oxidizing agent to oxidize the cytosine when it is methylated; and detecting the oxidized methylated cytosine.
摘要:
It is determined that the breast cancer has metastasized when methylation is detected in the CpG island of at least one selected from the group consisting of GSTP1, RASSF1A and RARb by detecting methylation in CpG island of each of GSTP1, RASSF1A and RARb contained in serum obtained from a patient who has undergone surgery to remove breast cancer.
摘要:
There is provided a method for confirming conversion treatment of non-methylated cytosine, including a step of mixing a DNA taken from a living body, with a nucleic acid for accuracy control having a sequence A which is not contained in a genome of the living body and contains cytosine, a step of converting non-methylated cytosine in the mixture into a base other than cytosine, a step of detecting the sequence A and/or a sequence A′ which has a sequence obtained by converting cytosine in the sequence A into the base other than cytosine, and a step of determining whether the conversion step has been properly performed or not based on the result of the detection step.
摘要:
The method for detecting a methylated cytosine of the present invention comprises the steps of:hybridizing a sample DNA with an oligonucleotide which can hybridize with a region of the sample DNA containing a cytosine suspected of being methylated and has an a basic site at the position complementary to the cytosine;reacting the hybridized sample DNA obtained in the previous step with an oxidizing agent to oxidize the cytosine when it is methylated; anddetecting the oxidized methylated cytosine.