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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.
Asian Pacific Journal of Cancer Prevention : APJCP
|July 29, 2026
Summary
A new, affordable molecular test for chronic myeloid leukemia (CML) was developed in Morocco. This reliable BCR-ABL1 detection assay improves access to essential CML monitoring for patients in resource-limited areas.
Area of Science:
- Molecular diagnostics
- Oncology
- Biotechnology
Background:
- Chronic myeloid leukemia (CML) is driven by the BCR-ABL1 fusion gene, necessitating molecular monitoring for effective patient management.
- Current imported molecular monitoring assays are costly, limiting access for CML patients in Morocco.
- Developing accessible diagnostic tools is crucial for improving healthcare outcomes in resource-limited settings.
Purpose of the Study:
- To develop and validate a cost-effective, one-step multiplex RT-qPCR assay for simultaneous detection of BCR-ABL1 and ABL1 transcripts.
- To establish a reliable and affordable alternative for routine BCR-ABL1 monitoring in Morocco.
- To enhance access to essential molecular testing for CML patients.
Main Methods:
- A one-step multiplex RT-qPCR assay was designed to detect BCR-ABL1 and ABL1 transcripts in peripheral blood mononuclear cells.
- The assay was validated using samples from 63 Moroccan CML patients.
- Performance was compared against the established GeneXpert system.
Main Results:
- The developed assay demonstrated high efficiency (>95%) and excellent linearity (R2 ≥ 0.99) for both BCR-ABL1 and ABL1 targets.
- BCR-ABL1/ABL1 ratios obtained from the new assay showed strong correlation with the GeneXpert system (r = 0.93, p < 0.001).
- An overall agreement of 86% was observed between the developed assay and the GeneXpert system.
Conclusions:
- The locally developed RT-qPCR assay is a dependable and economical option for routine BCR-ABL1 detection and monitoring in CML patients.
- This assay significantly improves access to molecular diagnostics in resource-limited settings like Morocco.
- The validated assay offers a practical solution for enhancing CML patient management through accessible molecular testing.
