Abstract: 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.

Abstract: 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.

Abstract: Provided are a primer set, a method and a kit therefor, which can easily perform with high accuracy the detection of a cancer cell caused by HPV and the determination of whether or not a tissue is a tissue with high-grade dysplasia or in a more severe phase. As a primer set, used is a primer set consisting of a first primer which hybridizes with a nucleic acid consisting of a nucleotide sequence in which cytosine present in a site other than a CpG site is converted into another base in a nucleotide sequence having a CpG site in L1 region or L2 region of HPV and a second primer which hybridizes with a nucleic acid consisting of a nucleotide sequence in which cytosine is converted into another base in a nucleotide sequence having a CpG site in LCR or E6 region of HPV.

Abstract: 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.

Abstract: The present invention relates to a method for determining presence or absence of a cancer cell in a biological sample based on the analysis result obtained by analyzing methylation status of DNA extracted from the biological sample with a novel molecular marker allowing a determination of presence or absence of the cancer cell.

Abstract: A method for detecting integrated HPV DNA is described herein. This method comprises obtaining first and second samples, obtaining first and second information, and detecting, based on the first and second information, the HPV DNA integrated into the genome of a cell derived from a subject. The second sample comprises DNA derived from the cell, which is treated with an enzyme having exonuclease activity. The first information is related to the amount of HPV DNA in the first sample, and the second information is related to the amount of HPV DNA in the second sample.

Abstract: In order to simply obtain a methylated DNA sample, a method including bringing a sample which may include methylated DNA into contact with a protein capable of binding to methylated DNA to bind the methylated DNA in the sample to the protein, bringing the sample obtained into contact with at least one deoxyribonuclease to degrade DNA in the sample; and detecting methylated DNA which is not degraded by the deoxyribonuclease by the binding of the sample obtained to the protein is used. In the method, at least one type of deoxyribonucleases is a deoxyribonuclease different from a methylation-sensitive restriction enzyme capable of degradating a single stranded DNA.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: A method for determining the presence or absence of an abnormal cell in a sample collected from the uterine cervix of a subject, and a method for predicting the progression of a lesion in the uterine cervix in a subject, each of which comprises measuring the frequency of methylation in the genomic DNA of human papillomavirus contained in the sample and determining the presence or absence of the abnormal cell or predicting the progression of the lesion based on the frequency; and a primer set which can be used in the above-mentioned methods.

Abstract: 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.

Abstract: 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.

Abstract: Provided are a primer set, a method and a kit therefor, which can easily perform with high accuracy the detection of a cancer cell caused by HPV and the determination of whether or not a tissue is a tissue with high-grade dysplasia or in a more severe phase. As a primer set, used is a primer set consisting of a first primer which hybridizes with a nucleic acid consisting of a nucleotide sequence in which cytosine present in a site other than a CpG site is converted into another base in a nucleotide sequence having a CpG site in L1 region or L2 region of HPV and a second primer which hybridizes with a nucleic acid consisting of a nucleotide sequence in which cytosine is converted into another base in a nucleotide sequence having a CpG site in LCR or E6 region of HPV.

Abstract: The invention provides a method of detecting, with a microarray, a target nucleic acid containing a base sequence to which a nucleic acid-binding protein binds, obtaining a signal from a correction probe not containing any sequence complementary to the base sequence to which the nucleic acid-binding protein binds for being used as a background to correct a signal obtained from a detection probe hybridizing to the target nucleic acid, as well as a computer program product for enabling a computer to detect, with a microarray, a target nucleic acid containing a base sequence to which a nucleic acid-binding protein binds.

Abstract: 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.

Abstract: 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.

Abstract: The present invention provides an easy and accurate method for obtaining information regarding a quantity of DNA included in a sample used for analyzing a methylation of a target DNA, comprising steps of: treating a biological sample containing DNA including the target DNA with a non-methylated cytosine converting agent for converting non-methylated cytosine of the DNA into another base; and obtaining information regarding a quantity of DNA included in the sample treated in the treating step by using an oligonucleotide complementary to a cytosine-free region of the target DNA.

Abstract: A method for detecting integrated HPV DNA is described herein. This method comprises obtaining first and second samples, obtaining first and second information, and detecting, based on the first and second information, the HPV DNA integrated into the genome of a cell derived from a subject. The second sample comprises DNA derived from the cell, which is treated with an enzyme having exonuclease activity. The first information is related to the amount of HPV DNA in the first sample, and the second information is related to the amount of HPV DNA in the second sample.

Abstract: 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.