Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay
| Autor: | László Rejtő, Péter Antal-Szalmás, Miklós Udvardy, László Szerafin, Zsuzsanna Hevessy, Renáta Hudák, János Kappelmayer, Valéria Kiss-Sziráki |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Clinical Biochemistry
Fusion Proteins bcr-abl Klinikai orvostudományok Polymerase Chain Reaction Peripheral blood mononuclear cell Flow cytometry hemic and lymphatic diseases medicine Humans Immunoassay Acute leukemia Leukemia biology medicine.diagnostic_test Chemistry Biochemistry (medical) Myeloid leukemia General Medicine Orvostudományok Reference Standards Flow Cytometry Molecular biology Fusion protein Microspheres Real-time polymerase chain reaction biology.protein Antibody Laboratories K562 cells |
| Popis: | A new flow cytometric (FC) BCR-ABL immunobead assay has been developed recently. Here we present the laboratory evaluation of the commercially available kit.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.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 (n=14) were positive with the FC method and negative results were also concordant (n=15). Frozen cell lysates can be stored up to 4 weeks without significant decrease of MFI signal.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. |
| Databáze: | OpenAIRE |
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