Source:http://linkedlifedata.com/resource/pubmed/id/10482989
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
9
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pubmed:dateCreated |
1999-9-30
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pubmed:abstractText |
To analyze the value of real time RT-PCR for monitoring of bcr-abl expression in CML patients after allogeneic or autologous stem cell transplantation (SCT), we generated pairs of PCR-primers and TaqMan probes specific for either the b2a2- or the b3a2-variant of bcr-abl. Either variant could be detected specifically from cDNA from a single K562 (b3a2) and BV173 (b2a2) cell with the respective TaqMan probe. Bcr-abl expression was normalized by comparison with GAPDH expression, and samples were quantitated using standard cDNA dilutions from K562 or BV173 cells. In a retrospective analysis 13 patients with CML after allogeneic (n = 10) or autologous (n = 3) SCT including patients with relapsed or persistent CML were analyzed by both real-time and conventional nested RT-PCR. In addition chimerism was monitored by FISH analysis of sex chromosomes in three patients with relapsed disease. The bcr-abl/GAPDH ratio dropped at least 1000-fold in all seven patients evaluable prior to and after allogeneic SCT as estimated by real-time RT-PCR, and conventional RT-PCR became negative in 6/7 patients. In five patients with relapsed or persistent disease after allogeneic SCT the bcr-abl/GAPDH ratio eventually increased again, and real-time RT-PCR was as sensitive as conventional RT-PCR for detection of bcr-abl. Donor lymphocyte infusions (DLI) were given to all five patients, and the bcr-abl/GAPDH ratio dropped to undetectable levels in two patients both remaining in continuing molecular remission. In contrast, in three other patients the bcr-abl/GAPDH ratio decreased only or did not change significantly after DLI. In three patients undergoing autologous SCT the bcr-abl/GAPDH ratio dropped only 1.1 to 30-fold, and the patients were tested positive with real-time RT-PCR at all time points. These data demonstrate that real-time RT-PCR is valuable to quantitate bcr-abl expression in CML patients after transplantation.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0887-6924
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
13
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1383-9
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:10482989-Bone Marrow Transplantation,
pubmed-meshheading:10482989-Computer Systems,
pubmed-meshheading:10482989-Female,
pubmed-meshheading:10482989-Fusion Proteins, bcr-abl,
pubmed-meshheading:10482989-Hematopoietic Stem Cell Transplantation,
pubmed-meshheading:10482989-Humans,
pubmed-meshheading:10482989-Leukemia, Myelogenous, Chronic, BCR-ABL Positive,
pubmed-meshheading:10482989-Male,
pubmed-meshheading:10482989-Retrospective Studies,
pubmed-meshheading:10482989-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:10482989-Transplantation, Autologous,
pubmed-meshheading:10482989-Transplantation, Homologous
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pubmed:year |
1999
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pubmed:articleTitle |
Monitoring of BCR-ABL expression using real-time RT-PCR in CML after bone marrow or peripheral blood stem cell transplantation.
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pubmed:affiliation |
Medizinische Hochschule Hannover, Abteilung Hämatologie und Onkologie, Zentrum der Inneren Medizin, Germany.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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