Patent ID: 9127317
Filing Date: 2015-09-08
Classification: C12Q

Abstract:
1. A method for monitoring beta cell death, comprising: extracting and purifying DNA from a body fluid of an animal or human; treating the extracted purified DNA with bisulfite to convert demethylated cytosine to uracil while sparing the methylated cytosines; amplifying the bisulfite-treated DNA using polymerase chain reaction; purifying the amplified bisulfite-treated DNA; providing methylation-status specific probes configured to measure, in a quantitative methylation-status sensitive probe hybridization reaction, demethylated insulin DNA of β cell origin and methylated insulin DNA of non-β cell origin, comprising: a) at least one probe for detection of methylated insulin DNA of non-β cell origin selected from the group consisting of ACCTCCCGACGAATCT SEQ ID NO: 003 and TACCTCTCGTCGAATCT SEQ ID NO: 004; and b) at least one probe for detection of demethylated insulin DNA of β cell origin selected from the group consisting of ACCTCCCAACAAATCT SEQ ID NO: 005 and TACCTCCCATCAAATCT SEQ ID NO: 006; performing the quantitative methylation sensitive probe hybridization reaction on the purified bisulfite-treated DNA using the methylation-status specific probes; and quantitatively analyzing relative amounts of methylated insulin DNA and demethylated insulin DNA.