title: Deep Sequencing of Cell-Free Peripheral Blood DNA as a Reliable Method for Confirming the Diagnosis of Myelodysplastic Syndrome.
abstract: BACKGROUND
  Demonstrating the presence of myelodysplastic syndrome (MDS)-specific molecular abnormalities can aid in diagnosis and patient management. We explored the potential of using peripheral blood (PB) cell-free DNA (cf-DNA) and next-generation sequencing (NGS).
  
  
  METHODS
  We performed NGS on a panel of 14 target genes using total nucleic acid extracted from the plasma of 16 patients, all of whom had confirmed diagnoses for early MDS with blasts <5%. PB cellular DNA from the same patients was sequenced using conventional Sanger sequencing and NGS.
  
  
  RESULTS
  Deep sequencing of the cf-DNA identified one or more mutated gene(s), confirming the diagnosis of MDS in all cases. Five samples (31%) showed abnormalities in cf-DNA by NGS that were not detected by Sanger sequencing on cellular PB DNA. NGS of PB cell DNA showed the same findings as those of cf-DNA in four of five patients, but failed to show a mutation in the RUNX1 gene that was detected in one patient's cf-DNA. Mutant allele frequency was significantly higher in cf-DNA compared with cellular DNA (p = 0.008).
  
  
  CONCLUSIONS
  These data suggest that cf-DNA when analyzed using NGS is a reliable approach for detecting molecular abnormalities in MDS and should be used to determine if bone marrow aspiration and biopsy are necessary.
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Ferras
authors.initials: F
authors.lastname: Albitar
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Wanlong
authors.initials: W
authors.lastname: Ma
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Kevin
authors.initials: K
authors.lastname: Diep
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Ivan
authors.initials: I
authors.lastname: De Dios
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Sally
authors.initials: S
authors.lastname: Agersborg
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Maya
authors.initials: M
authors.lastname: Thangavelu
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Steve
authors.initials: S
authors.lastname: Brodie
authors.affiliation: NeoGenomics Laboratories , Department of Hematology and Cytogenetics, Irvine, California.
authors.forename: Maher
authors.initials: M
authors.lastname: Albitar
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D004247:DNA / Q000097:blood* / Q000235:genetics; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009190:Myelodysplastic Syndromes / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D004247:DNA