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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

Online
ISSN
1437-4331
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Volume 47, Issue 4

Issues

Expression of lineage markers using real-time quantitative polymerase chain reaction (RT-qPCR) in normal and in leukemia bone marrow

Pascale Saussoy
  • Département de Biologie Clinique et d'Anatomie Pathologique, Cliniques Universitaires Saint-Luc (UCL), Bruxelles, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Jean-Luc Vaerman
  • Département de Biologie Clinique et d'Anatomie Pathologique, Cliniques Universitaires Saint-Luc (UCL), Bruxelles, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Vincent Druez
  • Département d'Orthopédie et de Traumatologie, Institut de Médecine, de Traumatologie et de Réadaptation (IMTR), Loverval, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Véronique Deneys / Nicole Straetmans / Guy Cornu / Augustin Ferrant
  • Département de Médecine Interne et Services Associés, Cliniques Universitaires Saint-Luc (UCL), Bruxelles, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Dominique Latinne
  • Département de Biologie Clinique et d'Anatomie Pathologique, Cliniques Universitaires Saint-Luc (UCL), Bruxelles, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar

Abstract

Background: The study of lineage markers by real-time quantitative polymerase chain reaction (RT-qPCR) at diagnosis enables differentiation between acute myeloblastic leukemia, B- or T-lineage acute lymphoblastic leukemia, without cell sorting. Our objective was to assess the relationship between protein expression and the amount of lineage marker mRNA in acute leukemia samples and to determine whether four lineage markers could be used to differentiate between normal and acute leukemia bone marrow (BM) without cell sorting.

Methods: Quantification of the mRNA of CD19, CD79a, CD3e, and myeloperoxidase was performed by RT-qPCR on 130 acute leukemia BM samples at diagnosis and on 20 BM samples from healthy donors, without cell sorting. Immunophenotyping of leukemia samples was performed after manual gating around the blastic population.

Results: Reference values for the four lineage markers were established by RT-qPCR for normal BM. The mRNA expression levels of these four lineage markers allowed the distinction between normal samples and 100% of acute leukemia samples.

Conclusions: With 92% congruence for protein expression and amount of mRNA in acute leukemias, these four lineage markers, essential for diagnosis and subclassification of acute leukemias by flow cytometry, also represent excellent candidate genes when using RT-qPCR technology as a diagnostic tool for molecular cancer class prediction.

Clin Chem Lab Med 2009;47:419–26.

Keywords: bone marrow; immunophenotyping; lineage marker; real-time PCR; reference values

About the article

Corresponding author: Dr. P. Saussoy, Laboratoire de Biologie Moléculaire, Tour R. Franklin, O Nord, Cliniques Universitaires Saint-Luc (UCL), avenue Hippocrate 10, 1200 Brussels, Belgium Phone: +32-2-7646780, Fax: +32-2-7646931,


Received: 2008-09-23

Accepted: 2009-01-05

Published in Print: 2009-04-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 47, Issue 4, Pages 419–426, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2009.093.

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[2]
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