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US 2010/0143893 A1Published application
Method for detection of cytosine methylation
OwnerKurt Berlininventor
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OwnerKurt Berlininventor
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Herein described is a method for the detection of cyto-sine methylation in a nucleic acid sample, comprising the steps of: a) treating a nucleic acid sample with an agent convert-ing unmethylated cytosine bases into uracil bases and not converting methylated cytosine bases within said nucleic acid sample, b) amplifying selected segments of the treated nucleic acid sample, by providing two first oligonucleotide primers (A and B) that are capable of producing an amplificate under certain chosen amplification conditions inde-pendently of the methylation status of the nucleic acid before treatment in step a), and further providing at least two additional second oligonucleotide primers (C and D) that can each produce a product with one of the first primers (A or B) under said same amplification conditions, wherein at least one of the second primers binds to the nucleic acid in a methylation specific manner, thereby distinguishing between unconverted initially methylated and converted unmethylated nucleic acids and/or blocking molecules are provided that hinder the binding of at least one of the second primers to the nucleic acid in a methylation specific manner, thereby distinguishing between unconverted initially methylated and converted unmethylated nucleic acids, c) detecting the amplificates of the treated nucleic acid.
A method for the detection of cytosine methylation in a nucleic acid sample, comprising the steps of:
A method according to one of the preceding claims, further characterized in that the presence of a disease or other medical condition of a patient is concluded from the degree of methylation at different CpG positions investigated.
A kit, comprising primers A, B, C and D, wherein primers A and B are capable of producing an amplificate from a bisulfite treated template nucleic acid sample, under suitable amplification conditions, and do not distinguish between prior to bisulfite treatment methylated and prior to bisulfite treatment unmethylated DNA,
A kit, comprising primers A, B, C and D, wherein primers A and B are capable of producing an amplificate from a bisulfite treated template nucleic acid sample, under suitable amplification conditions, and do not distinguish between prior to bisulfite treatment methylated and prior to bisulfite treatment unmethylated DNA,
Dependencies are inferred from the claim text.