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Standardizing minimal residual disease by flow cytometry for precursor B lineage acute lymphoblastic leukemia in a
Nikhil Patkar1, Ansu Abu Alex, Bargavi B
1Department of Hematology, Christian Medical College, Vellore, Tamil Nadu, India. nvpatkar@gmail.com
Background:
In addition to standard risk criteria at diagnosis, minimal residual disease (MRD) following initiation of therapy is a well-recognized risk factor to predict relapse. Literature from developing countries addressing therapeutic or laboratory practices related to MRD, is largely lacking. In a first paper from India, we describe our experience in establishing a flow cytometry-based MRD assay for precursor B lineage ALL (BCP-ALL) with emphasis on the assay standardization and cost.
Methods:
Normal templates for B cell development were established in 10 control patients using CD45, CD11a, CD38, CD20, CD10, CD19, CD58, CD34, CD123, and CD22. BCP-ALL samples (n = 42) were characterized at diagnosis to identify a suitable marker for follow-up during mid (D+21) and end of induction (D+33). Both, multiparametric immunophenotyping and single marker detection of LAIP were used for data analysis.
Results:
In 95.2% of BCP-ALL at least two informative markers could be obtained when a minimum of four cocktail combinations were used. The combination CD20, CD10, CD45, and CD19 was the most useful (71.4%) followed by combinations containing CD38 (66.7%), CD22 (57.1%), CD11a (52.4%), and CD58 (33.3%). Using our approach, 60 and 47% of patients had detectable MRD at mid and end induction time points, respectively.
Conclusion:
We have described a relatively cost effective MRD panel which is applicable to over 90% of patients. We hope that this data would encourage more centers in India and other resource constrained health delivery systems to develop MRD assays.
Insights
This study introduces a cost-effective flow cytometry assay for minimal residual disease (MRD) detection in precursor B-cell acute lymphoblastic leukemia (BCP-ALL). The assay is applicable to over 90% of patients, aiding relapse prediction in resource-limited settings.
Area of Science:
- Hematology
- Immunophenotyping
- Oncology
Background:
- Minimal residual disease (MRD) is a key predictor of relapse in B-cell acute lymphoblastic leukemia (ALL).
- Limited data exists on MRD assessment practices in developing countries.
- This study focuses on establishing a flow cytometry-based MRD assay for precursor B-cell ALL (BCP-ALL) in India.
Purpose of the Study:
- To describe the standardization and cost-effectiveness of a flow cytometry-based MRD assay for BCP-ALL.
- To identify suitable markers for MRD detection in BCP-ALL patients.
- To establish a reliable method for MRD assessment in a resource-constrained setting.
Main Methods:
- Established normal B cell development templates using a panel of 11 markers.
- Characterized 42 BCP-ALL samples at diagnosis to identify informative markers for MRD monitoring.
- Utilized multiparametric immunophenotyping and Last Appearance Immunophenotypic Marker (LAIP) detection for data analysis.
Main Results:
- At least two informative markers were identified in 95.2% of BCP-ALL cases using four-marker combinations.
- The combination of CD20, CD10, CD45, and CD19 was most effective (71.4%).
- MRD was detected in 60% and 47% of patients at mid and end of induction, respectively.
Conclusions:
- A cost-effective MRD panel was developed, applicable to over 90% of BCP-ALL patients.
- This assay can be implemented in resource-limited settings.
- Encourages the development of MRD assays in similar healthcare systems.
