epigenetic_qPCR_data_final.xlsx

We analysed whole blood and nasopharyngeal swabs from COVID-19 patients in two different cohorts collected at hospitals in Germany (Bochum) and Spain (Valencia) by epigenetic immune cell quantification using qPCR assays (demethyl-specific). The aim was to investigate the prognostic potential of this approach to identify patients with higher risk for a poor outcome. Also, we compared epigenetic data of patients with those of healthy donors. <br> Dataset includes Cp (crossing-point) values, cell specific plasmid units (PU) per qPCR reaction resulting from epigenetic qPCR analyses (calculated based on a quantification standard), as well as coefficient of variation (C.V.). PUs were translated into immune cell counts relative to the housekeeping gene GAPDH (glyceraldehyde 3-phosphate dehydrogenase) for each sample (peripheral whole blood, dried blood spots and swabs). GAPDH is also targeted by an epigenetic qPCR assay to quantify all GAPDH PUs. <br> All samples were lysed to extract genomic DNA. Afterwards, DNA was incubated with ammonium-bisulfite to convert all unmethylated CpGs (cytosin-guanine-dinucleotide) to uracil residues by deamination and desulphonation reaction without alteration of methylated CpGs. Within the subsequent epigenetic qPCR generated uracils are changed to thymine by proof-reading capability of the used DNA-Polymerase. <br> The epigenetic qPCR assays consist of demethyl-specific primer pairs and probes to detect cell type-specific demethylation within genes of interest. <br>