In:
Clinical Chemistry, Oxford University Press (OUP), Vol. 64, No. 3 ( 2018-03-01), p. 566-575
Abstract:
White blood cell counts are routinely measured with automated hematology analyzers, by flow cytometry, or by manual counting. Here, we introduce an alternative approach based on DNA methylation (DNAm) at individual CG dinucleotides (CpGs). METHODS We identified candidate CpGs that were nonmethylated in specific leukocyte subsets. DNAm levels (ranging from 0% to 100%) were analyzed by pyrosequencing and implemented into deconvolution algorithms to determine the relative composition of leukocytes. For absolute quantification of cell numbers, samples were supplemented with a nonmethylated reference DNA. RESULTS Conventional blood counts correlated with DNAm at individual CpGs for granulocytes (r = −0.91), lymphocytes (r = −0.91), monocytes (r = −0.74), natural killer (NK) cells (r = −0.30), T cells (r = −0.73), CD4+ T cells (r = −0.41), CD8+ T cells (r = −0.88), and B cells (r = −0.66). Combination of these DNAm measurements into the “Epi-Blood-Count” provided similar precision as conventional methods in various independent validation sets. The method was also applicable to blood samples that were stored at 4 °C for 7 days or at −20 °C for 3 months. Furthermore, absolute cell numbers could be determined in frozen blood samples upon addition of a reference DNA, and the results correlated with measurements of automated analyzers in fresh aliquots (r = 0.84). CONCLUSIONS White blood cell counts can be reliably determined by site-specific DNAm analysis. This approach is applicable to very small blood volumes and frozen samples, and it allows for more standardized and cost-effective analysis in clinical application.
Type of Medium:
Online Resource
ISSN:
0009-9147
,
1530-8561
DOI:
10.1373/clinchem.2017.279935
Language:
English
Publisher:
Oxford University Press (OUP)
Publication Date:
2018
