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.

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.

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. 

The epigenetic qPCR assays consist of demethyl-specific primer pairs and probes to detect cell type-specific demethylation within genes of interest.