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A machine learning framework for cross-institute standardized analysis of flow cytometry in differentiating acute myeloid leukemia from non-neoplastic conditions
Abstract
Flow cytometry (FC) remains a cornerstone diagnostic tool for acute myeloid leukemia (AML), yet standardizing panels across laboratories presents persistent challenges. Our study introduces a validated machine learning framework enabling cross-panel AML classification by leveraging common parameters shared across diverse FC protocols.
 
We employed FC data from 215 samples (110 AML, 105 non-neoplastic) collected in five institutions using different panel configurations as model training set, and another 196 similarly collected samples (90 AML and 106 non-neoplastic) for independent validation set. The framework employs GMM-SVM classification based on 16 common parameters (FSC-A, FSC-H, SSC-A, CD7, CD11b, CD13, CD14, CD16, CD19, CD33, CD34, CD45, CD56, CD64, CD117, and HLA-DR) that are consistently present across various panel designs. The framework demonstrated robust performance with 98.15 % accuracy, 99.82 % area under curve (AUC), 97.30 % sensitivity, and 99.05 % specificity. Independent validation on 196 additional samples further confirmed the framework's effectiveness, maintaining high performance with 93.88 % accuracy and 98.71 % AUC.
 
This research establishes the viability of standardized FC analysis across diverse panel configurations and instruments through machine learning implementation. The framework's robust performance suggests promising applications for harmonized multi-center FC analysis, potentially resolving current standardization challenges in flow cytometry interpretation.
Figures
Workflow of the presented panel-agnostic machine learning (ML) approach for AML versus non-nonneoplastic sample classification. This workflow is based on our previous sample classification method for a single panel. Common markers/parameters across different panels are selected and rearranged first before following cross-panel analysis.
Keywords
Flow cytometry | Acute myeloid leukemia | Machine learning | Universal algorithm
Publication Date
2025/06/01
Journal
Computers in Biology and Medicine
DOI
10.1016/j.compbiomed.2025.110394
Publisher
Elsevier