Comparison of Euroflow and Conventional Flow Cytometry Protocols for Minimal Residual Disease Detection in Pediatric B-Cell Acute Lymphoblastic Leukemia

Authors

Maldonado N, Katherine

Instituto Regional de Enfermedades Neoplásicas del sur – IREN SUR (Peru)

Mamani C, William

Instituto Regional de Enfermedades Neoplásicas del sur – IREN SUR (Peru)

Article Information

DOI: 10.47772/IJRISS.2025.91100134

Subject Category: Social science

Volume/Issue: 9/11 | Page No: 1657-1661

Publication Timeline

Submitted: 2025-11-10

Accepted: 2025-11-20

Published: 2025-12-02

Abstract

Acute Lymphoblastic Leukemia (ALL) is the most frequent hematological malignancy in childhood, and its prognosis is strongly linked to the monitoring of Minimal Residual Disease (MRD). Flow cytometry (FC) is the gold standard for MRD detection, but methods vary widely between laboratories, often leading to discrepancies in sensitivity and reliability. This study aimed to quantitatively and qualitatively compare two flow cytometry protocols—the standardized EuroFlow method and a conventional, laboratory-developed protocol—for MRD detection in pediatric B-cell ALL patients. A total of 228 bone marrow samples from 76 pediatric patients at IREN SUR were analyzed. The results, benchmarked against an external reference (INSNSB protocol), showed that 35.5% of patients were MRD-positive, predominantly in the female and older pediatric (11–15 years) subgroups. The EuroFlow protocol demonstrated a consistent 100% accuracy across all age and sex strata, whereas the conventional method showed diminished accuracy, falling to 83.3% in the female 11–15 years group. Crucially, both protocols maintained 100% specificity (precision). Correlation analysis indicated perfect diagnostic concordance for EuroFlow (kappa=1.000) and superior quantitative correlation with the reference for normal B-lymphoblasts (R=0.9798). These findings conclusively establish EuroFlow as the more reliable and robust protocol for MRD detection in a clinical setting, recommending its adoption to standardize diagnostic practice and improve relapse prediction.

Keywords

Acute lymphoblastic leukemia, minimal residual disease

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References

1. Aronoff, S., (1989). Geographic Information Systems: A Management Perspective. Ottawa: WDL Publications. [Google Scholar] [Crossref]

2. Álvarez-Zúñiga, C. D., Garza-Veloz, I., Martínez-Rendón, J., Ureño-Segura, M., Delgado-Enciso, I., & Martinez-Fierro, M. L. (2023). Circulating Biomarkers Associated with the Diagnosis and Prognosis of B-Cell Progenitor Acute Lymphoblastic Leukemia. Cancers, 15(16), 4186. https://doi.org/10.3390/cancers15164186. [Google Scholar] [Crossref]

3. Cossarizza, A., Chang, H. D., Radbruch, A., Acs, A., Adam, D., Adam-Klages, S., Agace, W. W., Aghaeepour, N., Akdis, M., Allez, M., Almeida, L. N., Alvisi, G., Anderson, G., Andrä, I., Annunziato, F., Anselmo, A., Bacher, P., Baldari, C. T., Bari, S. Zychlinsky, A. (2019). Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition). European Journal Of Immunology, 49(10), 1457-1973. https://doi.org/10.1002/eji.201970107. [Google Scholar] [Crossref]

4. De Smith, A. J., Jiménez-Morales, S., & Mejía-Aranguré, J. M. (2024). The genetic risk of acute lymphoblastic leukemia and its implications for children of Latin American origin. Frontiers In Oncology, 13. https://doi.org/10.3389/fonc.2023.1299355. [Google Scholar] [Crossref]

5. Ekpa, Q. L., Akahara, P. C., Anderson, A. M., Adekoya, O. O., Ajayi, O. O., Alabi, P. O., Okobi,, O. E., Jaiyeola, O., & Ekanem, M. S. (2023). A Review of Acute Lymphocytic Leukemia (ALL) in the Pediatric Population: Evaluating Current Trends and Changes in Guidelines in the Past Decade. Cureus, 15(12), e49930. https://doi.org/10.7759/cureus.49930. [Google Scholar] [Crossref]

6. Glier, H., Heijnen, I., Hauwel, M., Dirks, J., Quarroz, S., Lehmann, T., Rovo, A., Arn, K., Matthes, T., Hogan, C., Keller, P., Dudkiewicz, E., Stüssi, G., & Fernandez, P. (2019). Standardization of 8-color flow cytometry across different flow cytometer instruments: A feasibility study in clinical laboratories in Switzerland. Journal Of Immunological Methods, 475, 112348. https://doi.org/10.1016/j.jim.2017.07.013. [Google Scholar] [Crossref]

7. Juárez-Avendaño, G., Méndez-Ramírez, N., Luna-Silva, N. C., Gómez-Almaguer, D., Pelayo, R., & Balandrán, J. C. (2021). Molecular and cellular markers for measurable residual disease in acute lymphoblastic leukemia. Boletin médico del Hospital Infantil de México, 78(3), 159-170. https://doi.org/10.24875/BMHIM.20000155. [Google Scholar] [Crossref]

8. Kruse, A., Abdel-Azim, N., Kim, H. N., Ruan, Y., Phan, V., Ogana, H., Wang, W., Lee, R., Gang, E. J., Khazal, S., & Kim, Y. M. (2020). Minimal Residual Disease Detection in Acute Lymphoblastic Leukemia. International journal of molecular sciences, 21(3), 1054. https://doi.org/10.3390/ijms21031054. [Google Scholar] [Crossref]

9. Pieters, R., Mullighan, C. G., & Hunger, S. P. (2023). Advancing Diagnostics and Therapy to Reach Universal Cure in Childhood ALL. Journal of clinical oncology: official journal of the American Society of Clinical Oncology, 41(36), 5579-5591. https://doi.org/10.1200/JCO.23.01286. [Google Scholar] [Crossref]

10. Theunissen, P., Mejstrikova, E., Sedek, L., Van Der Sluijs-Gelling, A. J., Gaipa, G., Bartels, M., Da Costa, E. S., Kotrová, M., Novakova, M., Sonneveld, E., Buracchi, C., Bonaccorso, P., Oliveira, E., Marvelde, J. G. T., Szczepanski, T., Lhermitte, L., Hrusak, O., Lecrevisse, Q., Grigore, G. E.,... Van Der Velden, V. H. J. (2016). Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia. Blood, 129(3), 347-357. https://doi.org/10.1182/blood-2016-07-726307. [Google Scholar] [Crossref]

11. Van Dongen, J. J. M., Van Der Velden, V. H. J., Brüggemann, M., & Orfao, A. (2015). Minimal residual disease diagnostics in acute lymphoblastic leukemia: need for sensitive, fast, and standardized technologies. Blood, 125(26), 3996-4009. https://doi.org/10.1182/blood-2015-03-580027. [Google Scholar] [Crossref]

12. Verbeek, M. W. C., & van der Velden, V. H. J. (2024). The Evolving Landscape of Flowcytometric Minimal Residual Disease Monitoring in B-Cell Precursor Acute Lymphoblastic Leukemia. International journal of molecular sciences, 25(9), 4881. https://doi.org/10.3390/ijms25094881 [Google Scholar] [Crossref]

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