Real-time quantitation of bcr-abl transcripts in haematological malignancies.
We have applied an automated real-time quantitative PCR assay using a double-labeled fluorogenic probe to detect t(9;22)-positive cells in haematological malignancies. The results are expressed as the ratio of chimeric bcr-abl transcripts on abl transcripts. Highly reproducible results were obtained for t(9;22)-positive K562 RNA. Ten copies of bcr-abl DNA from a recombinant KW-3 plasmid and one positive cell in 10(4) can be detected. Thirty-two patients with chronic myeloid leukaemia (CML), 25 with acute leukaemia, 12 with myelodysplastic syndromes and 7 with other myeloproliferative syndromes were tested. Follow-up data were obtained in bcr-abl positive cases. Results were compared with those of conventional nested RT-PCR and cytogenetics. Real-time quantitative RT-PCR values correlated well with both these methods. However, in some cases the only means of detecting early relapse or blastic transformation was to examine the kinetics of real-time quantitative RT-PCR. Thus, real-time quantitative RT-PCR appears suitable for the diagnosis and follow-up of patients with the t(9;22) translocation.
Saffroy
R
R
Service de Biochimie et Biologie moléculaire-UPRES-EA 1596, Hôpital Paul Brousse and Faculté de Médecine Paris Sud Université Paris XI, Villejuif, France.
Lemoine
A
A
Brézillon
P
P
Frénoy
N
N
Delmas
B
B
Goldschmidt
E
E
Souleau
B
B
Nedellec
G
G
Debuire
B
B
D000208:Acute Disease; D001854:Bone Marrow Cells; D020732:Cytogenetic Analysis; D005260:Female; D005500:Follow-Up Studies; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D018628:Gene Dosage; D019337:Hematologic Neoplasms / Q000175:diagnosis / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis / Q000235:genetics; D007951:Leukemia, Myeloid / Q000175:diagnosis / Q000235:genetics; D007963:Leukocytes, Mononuclear; D008297:Male; D009190:Myelodysplastic Syndromes / Q000175:diagnosis / Q000235:genetics; D018365:Neoplasm, Residual / Q000175:diagnosis; D010677:Philadelphia Chromosome; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D012333:RNA, Messenger; D016044:Fusion Proteins, bcr-abl