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Development and Validation of a One-Step Multiplex RT-qPCR Assay for BCR-ABL1 Transcript Detection and Monitoring in
Abdenacer Benchrifa1,2,3, Basma Albanyahyati1,2,3, Mounyr Balouiri1
1Diagnostic Kits and Medical Device Development Center, Moroccan Foundation for Advanced Science, Innovation and Research (MAScIR), University Mohammed VI Polytechnic (UM6P), Ben Guerir, Morocco.
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.
