title: Application of allele-specific quantitative PCR using genomic DNA to monitor minimal residual disease based on mutant gene levels following allogeneic hematopoietic stem cell transplantation in patients with hematological malignancies: comparison of mutant levels with autologous DNA percentage by short tandem repeat-PCR.
abstract: BACKGROUND
  Quantitative evaluation of minimal residual disease (MRD) following hematopoietic stem cell transplantation (HSCT) is indispensable for patients with hematological malignancies. In addition to established MRD markers such as immunoglobulin and T-cell receptor gene rearrangements, fusion genes, or aberrantly expressed genes, single nucleotide mutations are considered one of the MRD markers that reflect the malignant cell clone.
  
  
  METHODS
  We compared the quantity of mutant genes by allele-specific quantitative polymerase chain reaction (AS-qPCR) for single nucleotide mutations (TP53 410T>A and PTPN11 1508G>A) with the percentage of autologous DNA by short tandem repeat (STR)-PCR.
  
  
  RESULTS
  Following HSCT, the quantity of mutant genes detected by AS-qPCR correlated with the percentage of autologous DNA assessed by the STR-PCR. Moreover, mutant DNAs were detected at a quantifiable level before relapse, whereas the percentage of autologous DNA was less than 5%, that is, complete chimerism.
  
  
  CONCLUSIONS
  The AS-qPCR approach for single nucleotide mutations was accurate and highly sensitive for monitoring pre-transplantation as well as post-transplantation MRD. AS-qPCR for single nucleotide mutation is suitable for monitoring MRD in patients who lack previously established MRD markers.
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Asahi, Matsumoto, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Taira
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsuda
authors.forename: Shoji
authors.initials: S
authors.lastname: Saito
authors.forename: Kazuo
authors.initials: K
authors.lastname: Sakashita
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Sugano
authors.forename: Nobuo
authors.initials: N
authors.lastname: Okumura
authors.forename: Takayuki
authors.initials: T
authors.lastname: Honda
mesh_terms: D000483:Alleles; D002675:Child, Preschool; D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis; D015894:Genome, Human / Q000235:genetics; D019337:Hematologic Neoplasms / Q000235:genetics / Q000473:pathology* / Q000601:surgery; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007223:Infant; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000473:pathology / Q000601:surgery; D057230:Limit of Detection; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics*; D009154:Mutation; D009190:Myelodysplastic Syndromes / Q000235:genetics / Q000473:pathology / Q000601:surgery; D018365:Neoplasm, Residual; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D014184:Transplantation, Homologous
chemical_list: D004247:DNA