PITX2 and breast carcinoma: The test utilizes real-time PCR with fluorescent hydrolysis probes and methylation-unspecific primers to detect methylation of three CpG sites within of PITX2 gene (assembly GRCh37/hg19, chr4: 111558429–111558431, chr4:111558435–111558437, chr4:111558443–111558445), in the DNA extracted from FFPE breast cancer tissue [89].