Quantification of BCR-ABL transcripts by QC-RT-PCR in leukapheresis-collected blood progenitor cells in early diagnosed patients with CML

M. T. Corsetti, F. Frassoni, F. Vassallo

Research output: Contribution to journalArticle

Abstract

The quality of the graft is thought to be essential for a good outcome in aulologuus stem cell transplant for CML. We have recently demonstrated that a) Ph-negative peripheral blood progenitor cells (PBPC) can be collected by leukapheresis during early recovery after chemotherapy-induced marrow aplasia, b) these cells are able to restore nORnal polyclonal hemopoiesis after autografting and c) the best clinical results are achieved in patients autografted with Ph-negative PBPC In order to monitor minimal residual disease in Ph-negative PBPC, we set a quantitative-competitive RT-PCR assay for BCR-ABL and analyzed Ph-negative collections in 18 patients mobilized in the first year from diagnosis without previous IFN-a treatment. We generated a construct m which the fusion region of the BCR-ABL gene was cloned in pCR-II vector and a 222 bp of foreign DNA was inserted in the ABL portion of the cDNA. This construct was used as competitor DNA to estimate BCR-ABL and ABL transcripts in 147 samples Ratio between the t\u> BCR-ABL and ABL was calculated for every sample. Statistical correlation between cytogenetics and QC-RT-PCR was higher than 0.8. Ph-negative collection from the same patients showed more than one log of difference in the amount of BCR-ABL transcripts, with the lower levels in the first two teukaphereses Sample BCR-ABL trans/ugRNA (median) BCR-ABL/ABL diagnosis 50000 0.3 1st leuk. 100 0.002 2nd leuk. 100 0.002 3rd leuk. 500 0.006 4th leuk 500 0.01 Sthleuk 1500 0.01 Indeed, the BCR-ABL level found in the first leukaphereses of these patients was lower than in the corresponding samples from a group of patients pre-treated for more than 12 months with IFN-α therapy. Patients mob BCR-ABL Irans/BCR-ABL/ABL agRNA fmedl in the 1 st leuk. (med) £12 mo. fromdx.inoINF 100 0.002 >12mo.IFN 1000 0.01 In conclusion, quantitative-competitive RT-PCR for BCR-ABL has been proven to be a powerful tool in monitoring leukemic contamination in PBPC collections. Follow-up data of patients autografted with low-contamination PBPCs are necessary to establish the clinical relevance of this selection.

Original languageEnglish
Pages (from-to)758
Number of pages1
JournalExperimental Hematology
Volume26
Issue number8
Publication statusPublished - 1998

Fingerprint

Leukapheresis
Blood Cells
Stem Cells
Polymerase Chain Reaction
Transplants
Autologous Transplantation
DNA
Residual Neoplasm
Cytogenetics
Complementary DNA
Bone Marrow
Drug Therapy
Therapeutics

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Quantification of BCR-ABL transcripts by QC-RT-PCR in leukapheresis-collected blood progenitor cells in early diagnosed patients with CML. / Corsetti, M. T.; Frassoni, F.; Vassallo, F.

In: Experimental Hematology, Vol. 26, No. 8, 1998, p. 758.

Research output: Contribution to journalArticle

@article{d15329404f634a80b186bf578363fb75,
title = "Quantification of BCR-ABL transcripts by QC-RT-PCR in leukapheresis-collected blood progenitor cells in early diagnosed patients with CML",
abstract = "The quality of the graft is thought to be essential for a good outcome in aulologuus stem cell transplant for CML. We have recently demonstrated that a) Ph-negative peripheral blood progenitor cells (PBPC) can be collected by leukapheresis during early recovery after chemotherapy-induced marrow aplasia, b) these cells are able to restore nORnal polyclonal hemopoiesis after autografting and c) the best clinical results are achieved in patients autografted with Ph-negative PBPC In order to monitor minimal residual disease in Ph-negative PBPC, we set a quantitative-competitive RT-PCR assay for BCR-ABL and analyzed Ph-negative collections in 18 patients mobilized in the first year from diagnosis without previous IFN-a treatment. We generated a construct m which the fusion region of the BCR-ABL gene was cloned in pCR-II vector and a 222 bp of foreign DNA was inserted in the ABL portion of the cDNA. This construct was used as competitor DNA to estimate BCR-ABL and ABL transcripts in 147 samples Ratio between the t\u> BCR-ABL and ABL was calculated for every sample. Statistical correlation between cytogenetics and QC-RT-PCR was higher than 0.8. Ph-negative collection from the same patients showed more than one log of difference in the amount of BCR-ABL transcripts, with the lower levels in the first two teukaphereses Sample BCR-ABL trans/ugRNA (median) BCR-ABL/ABL diagnosis 50000 0.3 1st leuk. 100 0.002 2nd leuk. 100 0.002 3rd leuk. 500 0.006 4th leuk 500 0.01 Sthleuk 1500 0.01 Indeed, the BCR-ABL level found in the first leukaphereses of these patients was lower than in the corresponding samples from a group of patients pre-treated for more than 12 months with IFN-α therapy. Patients mob BCR-ABL Irans/BCR-ABL/ABL agRNA fmedl in the 1 st leuk. (med) £12 mo. fromdx.inoINF 100 0.002 >12mo.IFN 1000 0.01 In conclusion, quantitative-competitive RT-PCR for BCR-ABL has been proven to be a powerful tool in monitoring leukemic contamination in PBPC collections. Follow-up data of patients autografted with low-contamination PBPCs are necessary to establish the clinical relevance of this selection.",
author = "Corsetti, {M. T.} and F. Frassoni and F. Vassallo",
year = "1998",
language = "English",
volume = "26",
pages = "758",
journal = "Experimental Hematology",
issn = "0301-472X",
publisher = "Elsevier Inc.",
number = "8",

}

TY - JOUR

T1 - Quantification of BCR-ABL transcripts by QC-RT-PCR in leukapheresis-collected blood progenitor cells in early diagnosed patients with CML

AU - Corsetti, M. T.

AU - Frassoni, F.

AU - Vassallo, F.

PY - 1998

Y1 - 1998

N2 - The quality of the graft is thought to be essential for a good outcome in aulologuus stem cell transplant for CML. We have recently demonstrated that a) Ph-negative peripheral blood progenitor cells (PBPC) can be collected by leukapheresis during early recovery after chemotherapy-induced marrow aplasia, b) these cells are able to restore nORnal polyclonal hemopoiesis after autografting and c) the best clinical results are achieved in patients autografted with Ph-negative PBPC In order to monitor minimal residual disease in Ph-negative PBPC, we set a quantitative-competitive RT-PCR assay for BCR-ABL and analyzed Ph-negative collections in 18 patients mobilized in the first year from diagnosis without previous IFN-a treatment. We generated a construct m which the fusion region of the BCR-ABL gene was cloned in pCR-II vector and a 222 bp of foreign DNA was inserted in the ABL portion of the cDNA. This construct was used as competitor DNA to estimate BCR-ABL and ABL transcripts in 147 samples Ratio between the t\u> BCR-ABL and ABL was calculated for every sample. Statistical correlation between cytogenetics and QC-RT-PCR was higher than 0.8. Ph-negative collection from the same patients showed more than one log of difference in the amount of BCR-ABL transcripts, with the lower levels in the first two teukaphereses Sample BCR-ABL trans/ugRNA (median) BCR-ABL/ABL diagnosis 50000 0.3 1st leuk. 100 0.002 2nd leuk. 100 0.002 3rd leuk. 500 0.006 4th leuk 500 0.01 Sthleuk 1500 0.01 Indeed, the BCR-ABL level found in the first leukaphereses of these patients was lower than in the corresponding samples from a group of patients pre-treated for more than 12 months with IFN-α therapy. Patients mob BCR-ABL Irans/BCR-ABL/ABL agRNA fmedl in the 1 st leuk. (med) £12 mo. fromdx.inoINF 100 0.002 >12mo.IFN 1000 0.01 In conclusion, quantitative-competitive RT-PCR for BCR-ABL has been proven to be a powerful tool in monitoring leukemic contamination in PBPC collections. Follow-up data of patients autografted with low-contamination PBPCs are necessary to establish the clinical relevance of this selection.

AB - The quality of the graft is thought to be essential for a good outcome in aulologuus stem cell transplant for CML. We have recently demonstrated that a) Ph-negative peripheral blood progenitor cells (PBPC) can be collected by leukapheresis during early recovery after chemotherapy-induced marrow aplasia, b) these cells are able to restore nORnal polyclonal hemopoiesis after autografting and c) the best clinical results are achieved in patients autografted with Ph-negative PBPC In order to monitor minimal residual disease in Ph-negative PBPC, we set a quantitative-competitive RT-PCR assay for BCR-ABL and analyzed Ph-negative collections in 18 patients mobilized in the first year from diagnosis without previous IFN-a treatment. We generated a construct m which the fusion region of the BCR-ABL gene was cloned in pCR-II vector and a 222 bp of foreign DNA was inserted in the ABL portion of the cDNA. This construct was used as competitor DNA to estimate BCR-ABL and ABL transcripts in 147 samples Ratio between the t\u> BCR-ABL and ABL was calculated for every sample. Statistical correlation between cytogenetics and QC-RT-PCR was higher than 0.8. Ph-negative collection from the same patients showed more than one log of difference in the amount of BCR-ABL transcripts, with the lower levels in the first two teukaphereses Sample BCR-ABL trans/ugRNA (median) BCR-ABL/ABL diagnosis 50000 0.3 1st leuk. 100 0.002 2nd leuk. 100 0.002 3rd leuk. 500 0.006 4th leuk 500 0.01 Sthleuk 1500 0.01 Indeed, the BCR-ABL level found in the first leukaphereses of these patients was lower than in the corresponding samples from a group of patients pre-treated for more than 12 months with IFN-α therapy. Patients mob BCR-ABL Irans/BCR-ABL/ABL agRNA fmedl in the 1 st leuk. (med) £12 mo. fromdx.inoINF 100 0.002 >12mo.IFN 1000 0.01 In conclusion, quantitative-competitive RT-PCR for BCR-ABL has been proven to be a powerful tool in monitoring leukemic contamination in PBPC collections. Follow-up data of patients autografted with low-contamination PBPCs are necessary to establish the clinical relevance of this selection.

UR - http://www.scopus.com/inward/record.url?scp=33748602916&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748602916&partnerID=8YFLogxK

M3 - Article

VL - 26

SP - 758

JO - Experimental Hematology

JF - Experimental Hematology

SN - 0301-472X

IS - 8

ER -