Microsphere-based multiplex analysis of DNA methylation in acute myeloid leukemia

Gerald B.W. Wertheim, Catherine Smith, Maria Figueroa, Michael Kalos, Adam Bagg, Martin Carroll, Stephen R. Master

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Aberrant regulation of DNA methylation is characteristic of cancer cells and clearly influences phenotypes of various malignancies. Despite clear correlations between DNA methylation and patient outcome, tests that directly measure multiple-locus DNA methylation are typically expensive and technically challenging. Previous studies have demonstrated that the prognosis of patients with acute myeloid leukemia can be predicted by the DNA methylation pattern of 18 loci. We have developed a novel strategy, termed microsphere HpaII tiny fragment enrichment by ligation-mediated PCR (MELP), to simultaneously analyze the DNA methylation pattern at these loci using methylation-specific DNA digestion, fluorescently labeled microspheres, and branched DNA hybridization. The method uses techniques that are inexpensive and easily performed in a molecular laboratory. MELP accurately reflects the methylation levels at each locus analyzed and segregates patients with acute myeloid leukemia into prognostic subgroups. Our results demonstrate the usefulness of MELP as a platform for simultaneous evaluation of DNA methylation of multiple loci.

Original languageEnglish (US)
Pages (from-to)207-215
Number of pages9
JournalJournal of Molecular Diagnostics
Volume16
Issue number2
DOIs
StatePublished - Mar 1 2014
Externally publishedYes

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DNA Methylation
Microspheres
Acute Myeloid Leukemia
Methylation
Ligation
Digestion
Neoplasms
Phenotype
Polymerase Chain Reaction
DNA

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Medicine

Cite this

Microsphere-based multiplex analysis of DNA methylation in acute myeloid leukemia. / Wertheim, Gerald B.W.; Smith, Catherine; Figueroa, Maria; Kalos, Michael; Bagg, Adam; Carroll, Martin; Master, Stephen R.

In: Journal of Molecular Diagnostics, Vol. 16, No. 2, 01.03.2014, p. 207-215.

Research output: Contribution to journalArticle

Wertheim, Gerald B.W. ; Smith, Catherine ; Figueroa, Maria ; Kalos, Michael ; Bagg, Adam ; Carroll, Martin ; Master, Stephen R. / Microsphere-based multiplex analysis of DNA methylation in acute myeloid leukemia. In: Journal of Molecular Diagnostics. 2014 ; Vol. 16, No. 2. pp. 207-215.
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