Patent ID: 7989169

Claim:
A method for amplification of a sample comprising a methylated target nucleic acid, and decontamination of the sample by removal of non-methylated contaminant target nucleic acid, the method consisting of the following steps in the sequence set forth: (a) performing a bisulfite reaction on the sample, thereby converting a non-methylated cytosine base into a uracil base while not converting a methylcytosine base, wherein the methylated target nucleic acid comprises at least one nucleotide with a methylcytosine base in a recognition nucleic acid sequence of a restriction enzyme; (b) adding a methylation-sensitive restriction enzyme to the sample which digests the non-methylated contaminant target nucleic acid, whereby after digestion the non-methylated contaminant target nucleic acid is no longer able to be amplified and the sample is decontaminated, whereby the methylated target nucleic acid is not digested by the methylation-sensitive restriction enzyme; (c) thermally inactivating the methylation-sensitive restriction enzyme; and (d) amplifying the methylated target nucleic acid, whereby after the amplifying step d) or concurrently with the amplifying step d), any amplification product produced in the amplifying step is detected as an indication of the presence or the amount of the methylated target nucleic acid in the sample.