The amount of BCR-ABL fusion transcripts detected by the real-time quantitative polymerase chain reaction method in patients with Philadelphia chromosome positive chronic myeloid leukemia correlates with the disease stage

The use of the real-time reverse-transcription polymerase-chain reaction (RT-PCR) method to quantify BCR-ABL transcripts before and after allogeneic transplant was prospectively studied in 65 patients with chronic myeloid leukemia (CML). The expression of the BCR-ABL transcript was determined and no...

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Bibliographic Details
Published in:Annals of hematology 2000-08, Vol.79 (8), p.424-431
Main Authors: ELMAAGACLI, A. H, BEELEN, D. W, OPALKA, B, SEEBER, S, SCHAEFER, U. W
Format: Article
Language:English
Subjects:
Abl protein
alpha -Interferon
BCR protein
Biological and medical sciences
Blast crisis
Bone Marrow Transplantation
Chronic myeloid leukemia
Donors
Fusion protein
Fusion Proteins, bcr-abl - genetics
Glyceraldehyde-3-phosphate dehydrogenase
Hematologic and hematopoietic diseases
Hematopoietic Stem Cell Transplantation
Humans
Immunosuppression
Immunotherapy
Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics
Leukemia, Myeloid, Chronic-Phase - genetics
Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis
Leukocytes
Medical sciences
Neoplasm Recurrence, Local
Philadelphia chromosome
Polymerase chain reaction
Recurrence
Reverse Transcriptase Polymerase Chain Reaction - methods
RNA, Messenger - analysis
Sensitivity and Specificity
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Summary:The use of the real-time reverse-transcription polymerase-chain reaction (RT-PCR) method to quantify BCR-ABL transcripts before and after allogeneic transplant was prospectively studied in 65 patients with chronic myeloid leukemia (CML). The expression of the BCR-ABL transcript was determined and normalized using the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeping gene product as an endogenous reference. In the single step real-time PCR assay, tenfold serial dilutions of cDNA of the K5652 cell line remained positive down to 100 pg cDNA only. However, molecular relapses of CML after transplant were only safely detectable when a nested real-time PCR assay was performed, which was able to detect 1-10 pg cDNA from a tenfold serial dilution. The median normalized BCR-ABL transcript level was measured as 0.004% in 17 patients with a molecular relapse, 0.4% in 7 patients with a cytogenetic relapse, 2.6% in 36 patients with a stable phase of CML, and 36% in 5 patients with a relapse in a blast crisis. The analyzed median normalized amount of BCR-ABL transcript differed significantly (P
ISSN:0939-5555
1432-0584
DOI:10.1007/s002770000169