Method for epigenetic immune cell detection and counting in human blood samples

A method for producing and detecting a panel of amplicons, the method comprising the steps of

• a) bisulfite treating isolated genomic DNA from a human blood cell sample containing immune cells to convert unmethylated cytosines into uracil;
• b) amplifying the bisulfite treated DNA to produce the panel of amplicons;
• wherein said amplifying comprises amplification and/or qPCR using:
• a thymidine-cytosine (TpG) template carrying targets of converted genomic regions of CD3gamma/delta, CD4, CD8beta, lipoprotein receptor-related protein 5 (LRP5), mevalonate pyrophosphate decarboxylase (MVD), and lipocalin-2 (LCN2), and
• primer pairs and probes for the converted genomic regions of CD3gamma/delta, CD4, CD8beta, LRP5, MVD, and LCN2, the primer pairs and probes selected from SEQ ID NOs:
• 3 to 12 for CD4, SEQ ID NOs:
• 13 to 20 for CD8beta, SEQ ID NOs:
• 21 to 28 for LRP5, SEQ ID NOs:
• 29 to 36 for MVD, SEQ ID NOs:
• 37 to 44 for LCN2, and SEQ ID NOs:
• 45 to 56 for CD3gamma/delta, and
• c) detecting the panel of amplicons.