These authors contributed equally to this work.
Detecting minimal residual disease in patients with chronic lymphocytic leukemia using 8-color flow cytometry protocol in routine hematological practice
Article first published online: 13 SEP 2013
© 2013 John Wiley & Sons Ltd
International Journal of Laboratory Hematology
Volume 36, Issue 2, pages 165–171, April 2014
How to Cite
Stehlíková, O., Chovancová, J., Tichý, B., Krejčí, M., Brychtová, Y., Panovská, A., Francová Skuhrová, H., Burčková, K., Borský, M., Loja, T., Mayer, J., Pospíšilová, Š. and Doubek, M. (2014), Detecting minimal residual disease in patients with chronic lymphocytic leukemia using 8-color flow cytometry protocol in routine hematological practice. International Journal of Laboratory Hematology, 36: 165–171. doi: 10.1111/ijlh.12149
- Issue published online: 15 MAR 2014
- Article first published online: 13 SEP 2013
- Manuscript Accepted: 16 JUL 2013
- Manuscript Received: 27 MAR 2013
- CEITEC. Grant Number: MSMT CR CZ.1.05/1.1.00/02.0068
- SuPReMMe. Grant Numbers: MSMT CR CZ.1.07/2.3.00/20.0045, MSM0021622430
- European Commission under the Health Theme of the 7th Framework Programme for Research and Technological Development. Grant Number: GA 306242
- Internal Grant Agency, Ministry of Health. Grant Number: NT13493-4/2012
- Czech Leukemia Study Group – for Life. Grant Numbers: MUNI/A/0723/2012, MH CZ - DRO (FNBr. 65269705)
- Chronic lymphocytic leukemia;
- flow cytometry;
- minimal residual disease
Minimal residual disease (MRD) detection has become increasingly important for the assessment of therapy response in chronic lymphocytic leukemia (CLL). However, current MRD analysis methods, both molecular genetic and flow cytometric, are time-consuming and require experienced laboratory staff.
To reduce the demands of flow cytometric MRD detection in CLL, we have introduced a novel flow cytometric 8-color protocol. The MRD analysis results using this protocol were then compared with the commonly employed 4-color protocol and the molecular genetic (real-time quantitative allele-specific oligonucleotide IGH polymerase chain reaction; RQ-ASO IGH PCR) approach.
Forty-two CLL patient samples were repeatedly analyzed after allogeneic stem cell transplantation (n = 20) or after fludarabine-based therapy (n = 22), and 100% concordance was found using both flow cytometric protocols. Furthermore, there was a strong correlation (r = 0.94) between flow cytometric and RQ-ASO IGH PCR results in MRD detection.
Flow cytometry is less time-consuming, less financially demanding, and moreover, MRD assessment using our novel 8-color protocol is less complicated than the 4-color approach and molecular methods.