title: Microsphere-based multiplex analysis of DNA methylation in acute myeloid leukemia.
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.
authors.affiliation: Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania; Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Gerald B W
authors.initials: GB
authors.lastname: Wertheim
authors.affiliation: Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania.
authors.forename: Catherine
authors.initials: C
authors.lastname: Smith
authors.affiliation: Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Figueroa
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania; Abramson Research Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Michael
authors.initials: M
authors.lastname: Kalos
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Adam
authors.initials: A
authors.lastname: Bagg
authors.affiliation: Division of Hematology and Oncology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Martin
authors.initials: M
authors.lastname: Carroll
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. Electronic address: srmaster@mail.med.upenn.edu.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Master
mesh_terms: D019175:DNA Methylation; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis / Q000235:genetics*; D008863:Microspheres; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D011379:Prognosis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity