ZAP-70 intron1 DNA methylation status: determination by pyrosequencing in B chronic lymphocytic leukemia

Leuk Res. 2010 Jun;34(6):800-8. doi: 10.1016/j.leukres.2009.10.018. Epub 2009 Nov 26.


ZAP-70 expression is a strong prognostic indicator in chronic lymphocytic leukemia. However, ZAP-70 quantification by flow cytometry lacks sufficient standardization. Based upon the correlation between ZAP-70 expression and its gene methylation status, we have developed a quantitative pyrosequencing assay for the determination of ZAP-70 methylation adapted for routine use. Methylation in four CpG pairs (C-223, C-243, C-254, and C-267) in the first intron of ZAP-70 is associated with repression of ZAP-70. Moreover, it correlates with CD38 expression (n=111, p<.0001), IgHv mutation status (n=106, p<.0001), time to treatment (p<.0001), and overall survival (p=.0014). Pyrosequencing of ZAP-70 provides a good alternative to flow cytometry.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • DNA Methylation*
  • Disease Progression
  • Female
  • Flow Cytometry / methods
  • Gene Expression Regulation, Leukemic
  • Humans
  • Introns / genetics
  • Leukemia, Lymphocytic, Chronic, B-Cell / diagnosis
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics*
  • Leukemia, Lymphocytic, Chronic, B-Cell / pathology
  • Male
  • Middle Aged
  • Predictive Value of Tests
  • Prognosis
  • Sequence Analysis, DNA / methods*
  • ZAP-70 Protein-Tyrosine Kinase / genetics*


  • ZAP-70 Protein-Tyrosine Kinase
  • ZAP70 protein, human