title: Molecular diagnosis of Philadelphia chromosome-positive chronic myeloid leukemia.
abstract: BACKGROUND
  In virtually all Ph1 chromosome-positive CML patients, the breakpoint on chromosome 22 maps in a very restricted area of 5.8 Kb, which has been named "breakpoint cluster region" or "bcr". Several molecular probes of this region are presently available, and this makes the molecular diagnosis of CML a useful approach which can be particularly important in those cases in which cytogenetic analysis does not reveal the presence of a Ph1 chromosome. Here we report the problems and our experience during the molecular analysis of the 478 patients examined so far.
  
  
  METHODS
  Molecular analyses were performed after digestion of the DNA with 2 to 4 restriction enzymes and hybridization with different probes. Individual samples were subjected to PCR since no rearrangements had been obtained with Southern blotting.
  
  
  RESULTS
  Rearrangement bands were detected in all the samples examined. In 473 cases the breakpoint was located within the bcr. In one of these cases, it was detected only after PCR analysis, and in two cases only after the use of the PHL/BCR probe. In 5 cases the breakpoint was localized either 5' or 3' with respect to the bcr.
  
  
  CONCLUSIONS
  In this paper, the criteria for a correct molecular diagnosis of CML are presented. The "PHL/BCR" probe appeared to be very specific and time-saving, since it required only one digestion to evidence the rearrangement. Our results confirm the high specificity of the breakpoint on chromosome 22 in CML and the relatively rare incidence of molecular variants.
authors.affiliation: Istituto di Ematologia Lorenzo e Ariosto Seràgnoli, Università di Bologna, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Zaccaria
authors.forename: A
authors.initials: A
authors.lastname: Tassinari
authors.forename: A
authors.initials: A
authors.lastname: Guerrasio
authors.forename: C
authors.initials: C
authors.lastname: Rosso
authors.forename: F
authors.initials: F
authors.lastname: Lo Coco
authors.forename: D
authors.initials: D
authors.lastname: Diverio
authors.forename: N
authors.initials: N
authors.lastname: Testoni
authors.forename: B
authors.initials: B
authors.lastname: Celso
authors.forename: M
authors.initials: M
authors.lastname: Buzzi
authors.forename: S
authors.initials: S
authors.lastname: Tura
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D004262:DNA Restriction Enzymes; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D015321:Gene Rearrangement; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010677:Philadelphia Chromosome; D016133:Polymerase Chain Reaction
chemical_list: D015345:Oligonucleotide Probes; D016044:Fusion Proteins, bcr-abl; D004262:DNA Restriction Enzymes