| CPC C12Q 1/6886 (2013.01) [C12Q 2600/106 (2013.01); C12Q 2600/118 (2013.01); C12Q 2600/154 (2013.01)] | 6 Claims |
|
1. A method for determining an increased risk of breast cancer in a human subject, wherein the method comprises:
(a) providing (i) a biological sample containing DNA from the human subject and extracting cell-free DNA from the sample or (ii) a biological sample from a breast tumor tissue of the human subject and extracting the genomic DNA from the sample, wherein the cell-free DNA or the genomic DNA comprises the targets of GCM2 and ITPRIPL1 DNA;
(b) assaying methylation level of the GCM2 and ITPRIPL1 from the cell-free DNA sample or the genomic DNA sample using methylation specific polymerase chain reaction assay (MSP), wherein the forward and reverse primers used in the assay comprise SEQ ID NO: 5-6 for GCM2 and SEQ ID NO: 7-8 for ITPRIPL1:
GCM2 forward primer: GAGATAGGGCGGAGTTTTTC (SEQ ID NO:5)
GCM2 reverse primer: CTTAACCGCGATACTAAACGTT (SEQ ID NO:6)
ITPRIPL1 forward primer: GAGTGTAGTTGATAGTAGGTACGGC (SEQ ID NO:7)
ITPRIPL1 reverse primer: GTAAATTTACTAAAAAAATAAAAAAACCGT (SEQ ID NO:8);
(c) comparing the methylation levels of Step (b) to methylation levels from a control sample without breast cancer; and
(d) determining an increased risk of breast cancer in the human subject based upon a higher methylation level detected in the sample as compared to the control sample.
|