Development and Validation of a One-Step Multiplex RT-qPCR Assay for BCR-ABL1 Transcript Detection and Monitoring in Moroccan Chronic Myeloid Leukemia Patients

Benchrifa, Abdenacer; Albanyahyati, Basma; Balouiri, Mounyr; El Hadi, Hicham; Bakri, Youssef; Doghmi, Kamal; Moumen, Abdeladim

doi:10.31557/APJCP.2026.27.7.2481

Development and Validation of a One-Step Multiplex RT-qPCR Assay for BCR-ABL1 Transcript Detection and Monitoring in Moroccan Chronic Myeloid Leukemia Patients

Document Type : Research Articles

Authors

¹ Diagnostic Kits and Medical Device Development Center, Moroccan Foundation for Advanced Science, Innovation and Research (MAScIR), University Mohammed VI Polytechnic (UM6P), Ben Guerir, Morocco.

² Laboratory of Human Pathologies Biology, Department of Biology, Faculty of Sciences, Morocco.

³ Genomic Center of Human Pathologies, Faculty of Medicine and Pharmacy, University Mohammed V, Rabat, Morocco.

⁴ Department of Clinical Hematology, Military Hospital of Instruction Mohamed V, Rabat, Morocco.

10.31557/APJCP.2026.27.7.2481

Abstract

Objective: Chronic myeloid leukemia (CML) is driven by the BCR-ABL1 fusion gene. Molecular monitoring of this transcript is essential for patient management. However, the high cost of imported assays limits access to regular monitoring in Morocco. Methods: We developed and validated a cost-effective one-step multiplex RT-qPCR assay that simultaneously detects BCR-ABL1 and ABL1 transcripts in peripheral blood mononuclear cells. Samples from 63 Moroccan CML patients were analyzed and compared with the GeneXpert system. Result: The assay showed efficiencies above 95% and R2 ≥ 0.99 for both targets. BCR-ABL1/ABL1 ratios correlated strongly with GeneXpert (r = 0.93, p < 0.001) and showed 86% overall agreement. Conclusion: This locally developed assay is a reliable and affordable alternative for routine BCR-ABL1 detection and monitoring, improving access to molecular testing in resource-limited settings.

Keywords