title: Molecular monitoring of tumour load kinetics predicts disease progression after non-myeloablative allogeneic stem cell transplantation in multiple myeloma.
abstract: BACKGROUND
  Non-myeloablative allogeneic stem cell transplantation followed by immunomodulatory therapies is considered a potentially curative approach in the treatment of multiple myeloma and most effective in a minimal residual disease setting.
  
  
  METHODS
  The aim of this study was to find the most sensitive real-time PCR assay (TaqMan), based on the IGH rearrangement, to quantify the tumour load of 11 patients with multiple myeloma after non-myeloablative allogeneic transplantation. Patient-allele specific primers (ASO) and the TaqMan probe were derived from CDR2 and CDR3 hypervariable regions of IGH, while consensus primers were located within the FR3 and FR4/JH regions. Four different approaches of primer combinations were tested.
  
  
  RESULTS
  ASO-forward and -reverse primers together with the clone-specific TaqMan probe were the most sensitive approach compared with the JH (P=0.071) or the FR3 consensus primer (P <0.001). The detection limit amounted to 1/10(4)-1/10(5) cells. Consecutively, 120 samples from 11 patients prior and post allogeneic transplantation were analysed. Three patients reached complete clinical remission accompanied by molecular remission. Disease progression or relapse was seen in six patients. In five, molecular progressive disease was detected prior to the clinical diagnosis of progression or relapse.
  
  
  CONCLUSIONS
  Patient-specific real-time IGH-PCR provides the opportunity for earlier treatment intervention.
authors.affiliation: Department of Internal Medicine V, University of Heidelberg, Germany. marc.raab@med.uni-heidelberg.de
authors.forename: M S
authors.initials: MS
authors.lastname: Raab
authors.forename: F W
authors.initials: FW
authors.lastname: Cremer
authors.forename: I N
authors.initials: IN
authors.lastname: Breitkreutz
authors.forename: S
authors.initials: S
authors.lastname: Gerull
authors.forename: T
authors.initials: T
authors.lastname: Luft
authors.forename: A
authors.initials: A
authors.lastname: Benner
authors.forename: M
authors.initials: M
authors.lastname: Goerner
authors.forename: A D
authors.initials: AD
authors.lastname: Ho
authors.forename: H
authors.initials: H
authors.lastname: Goldschmidt
authors.forename: M
authors.initials: M
authors.lastname: Moos
mesh_terms: D000328:Adult; D018450:Disease Progression; D006801:Humans; D008875:Middle Aged; D015166:Monitoring, Immunologic / Q000379:methods; D009101:Multiple Myeloma / Q000276:immunology / Q000473:pathology* / Q000628:therapy*; D011237:Predictive Value of Tests; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D033581:Stem Cell Transplantation / Q000379:methods*; D014184:Transplantation, Homologous; D047368:Tumor Burden / Q000276:immunology