Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay

Zsuzsanna Hevessy, Renta Hudk, Valria Kiss-Szirki, P. Antal-Szalmás, M. Udvardy, Lszl Rejt, Lszl Szerafin, J. Kappelmayer

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background: A new flow cytometric (FC) BCR-ABL immunobead assay has been developed recently. Here we present the laboratory evaluation of the commercially available kit. Methods: Mononuclear cells were isolated, lysed and processed according to the instructions of the manufacturer. Anti-BCR antibodies adsorbed to capture beads bind the BCR-ABL fusion proteins of the lysed cells, a phycoerythrin (PE)-conjugated anti-ABL antibody is the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Detection of t(9;22)(q34;q11) translocation was carried out with a quantitative PCR assay. Results: MFI results of 20 normal peripheral blood samples were 88±8 (mean±SD), CV 9. K562 cells were used as positive control. Within-batch imprecision was excellent (3.7 in the normal and 10 in the pathological range). Cut-off was chosen at MFI 112, where both sensitivity and specificity were 100. Altogether 17 chronic myeloid leukemia (CML) and 16 acute leukemia samples were analyzed. All PCR positive samples (n14) were positive with the FC method and negative results were also concordant (n15). Frozen cell lysates can be stored up to 4 weeks without significant decrease of MFI signal. Conclusions: The FC BCR-ABL assay is a fast, reproducible and reliable method that may be incorporated into standard flow cytometric protocols to help clinical decision-making.

Original languageEnglish
Pages (from-to)689-692
Number of pages4
JournalClinical Chemistry and Laboratory Medicine
Volume50
Issue number4
DOIs
Publication statusPublished - Apr 1 2012

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Assays
Fluorescence
Anti-Idiotypic Antibodies
Phycoerythrin
Polymerase Chain Reaction
K562 Cells
Flow cytometry
Antibodies
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Flow Cytometry
Leukemia
Blood
Fusion reactions
Decision making
Detectors
Sensitivity and Specificity
Proteins

Keywords

  • BCR-ABL
  • flow cytometry
  • immunobead assay

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay. / Hevessy, Zsuzsanna; Hudk, Renta; Kiss-Szirki, Valria; Antal-Szalmás, P.; Udvardy, M.; Rejt, Lszl; Szerafin, Lszl; Kappelmayer, J.

In: Clinical Chemistry and Laboratory Medicine, Vol. 50, No. 4, 01.04.2012, p. 689-692.

Research output: Contribution to journalArticle

Hevessy, Zsuzsanna ; Hudk, Renta ; Kiss-Szirki, Valria ; Antal-Szalmás, P. ; Udvardy, M. ; Rejt, Lszl ; Szerafin, Lszl ; Kappelmayer, J. / Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay. In: Clinical Chemistry and Laboratory Medicine. 2012 ; Vol. 50, No. 4. pp. 689-692.
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