MRD Detection Development Services for CML

The BCR-ABL1 fusion gene serves as a crucial marker for chronic myeloid leukemia (CML) diagnosis and the monitoring of minimal residual disease (MRD). Alfa Cytology has developed a diverse range of technologies for the detection of BCR-ABL1, enabling the identification of residual leukemia cells to evaluate treatment effectiveness and predict the likelihood of relapse. Our comprehensive services encompass the development of methods for monitoring CML MRD, supporting early-stage research in CML treatment, and providing effective solutions for preclinical studies.

Introduction to MRN in CML

The successful utilization of tyrosine kinase inhibitors (TKIs) has revolutionized the treatment of chronic myeloid leukemia (CML), resulting in a complete hematological response within approximately three months of initiating therapy. The detection of BCR-ABL1 transcript levels using biological techniques and methods is the established approach for monitoring minimal residual disease (MRD) in CML. The regular and standardized monitoring of MRD in CML plays a vital role in determining treatment response, selecting the optimal therapeutic approach, and providing valuable prognostic information.

Fig. 1. Measurable residual disease in chronic myeloid leukemia can be measured using RQ-PCR or ddPCR. (Branford, S.; Apperley, J.F., 2022)

The main strategy of our development services is based on the detection of BCR-ABL1 fusion genes, transcripts, or proteins. Additionally, we have devised alternative approaches aimed at developing innovative detection methods, with a focus on standardizing CML MRD monitoring.

Development of BCR-ABL1-Dependent MRD Detection

Molecular-based Approaches

• We offer an RQ-PCR method for molecular monitoring of CML MRD, based on specific primer and probe systems designed for the BCR and ABL1 genes. We have developed novel reference genes, such as ABL1 and GUSB, for detection purposes. Due to the variability and heterogeneity that can arise from numerous experimental steps and technical details in RQ-PCR analysis, we have also developed standardized experimental procedures to enhance the reliability of the method.
• We have developed a novel sensitive system based on long-range PCR amplification and deep sequencing to identify variants or mutations in BCR-ABL1. By comparing the BCR-ABL1 transcript levels with the standardized IS baseline, the outcomes of CML following TKI treatment can be evaluated.
• By developing digital droplet PCR (ddPCR) for monitoring BCR-ABL1 transcripts, we enable molecular monitoring of CML MRD. The dPCR method we have developed offers higher accuracy and sensitivity in measuring specific nucleic acid abundances, allowing for absolute quantification and providing greater precision and reproducibility compared to RQ-PCR.
• The multiple RQ-PCR method we have developed allows for the high sensitivity and specificity detection of various BCR-ABL1 fusion gene subtypes in a single reaction, enabling precise MRD monitoring. This approach addresses the limitation of most RQ-PCR methods that can only detect a limited number of BCR-ABL1 isoforms at a time.
• Based on sanger sequencing and next-generation testing, we have developed multiple strategies to characterize the specific BCR-ABL1 breakpoints in CML.
• We have developed various methods to test for the presence of BCR-ABL1 in the genome, including qPCR, RQ-PCR, RT-dPCR, and ddPCR. If your goal is to develop the most sensitive method, we recommend ddPCR.

Protein-Based Approaches

We have developed flow cytometry assays for the detection of BCR-ABL1 fusion proteins (p190, p210, and p230) in cell lysates, enabling CML MRD monitoring. Additionally, we offer method development for other techniques that combine flow cytometry with detection assays, such as PLA-flow. This approach exhibits high sensitivity to detect low-abundance protein targets within cell populations.

Development of BCR-ABL1-Independent MRD Detection

Due to the limitations of BCR-ABL1-based MRD detection, we offer method development in the following aspects.

Advantages of Our Services

• We utilize molecular biology and genomics to develop highly sensitive and specific assays for MRD detection.
• We offer personalized MRD monitoring services tailored to scientific research.
• We provide timely reporting of MRD detection results, accompanied by comprehensive data analysis.
• We offer innovative approach development services for MRD detection in CML.

Alfa Cytology is dedicated to advancing the precision and effectiveness of CML management. Partner with us to harness the power of MRD detection and make informed decisions in the fight against CML. Contact us for further information regarding our services in the development of MRD detection for CML.