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Simultaneous Quantification of T-Cell Receptor Excision Circles (TRECs) and K-Deleting Recombination Excision Circles (KRECs) by Real-time PCR
Published on: December 6, 2014
Quantification of minimal residual disease in T-lineage acute lymphoblastic leukemia with the TAL-1 deletion using a
1Department of Hematology/Oncology, St Jude Children's Research Hospital, Memphis, TN 38105, USA.
Abstract:
Hematologic relapse remains the greatest obstacle to the cure of children with acute lymphoblastic leukemia (ALL). Recent studies have shown that patients with increased risk of relapse can be identified by measuring residual leukemic cells, called minimal residual disease (MRD), during clinical remission. Current PCR methods, however, for measuring MRD are cumbersome and time-consuming. To improve and simplify MRD assessment, we developed a real-time quantitative PCR (RQ-PCR) assay for detection of leukemic cells that harbor the TAL-1 deletion. We studied serial dilutions of leukemic DNA and found the assay had a sensitivity of detection of one leukemic cell among 100,000 normal cells. We then investigated 23 samples from eight children with ALL in clinical remission. We quantified residual leukemic cells by using the TAL-1 RQ-PCR assay and by using limiting dilution analysis. In 17 samples, both methods detected MRD levels > or =0.001%. The percentages of leukemic cells measured by the two methods correlated well (r2 = 0.926). In the remaining six samples, both methods detected fewer than 0.001% leukemic cells. We conclude the TAL-1 RQ-PCR assay can be used for rapid, sensitive and accurate assessment of MRD in T-lineage ALL with the TAL-1 deletion.
Insights
A new real-time quantitative PCR (RQ-PCR) assay accurately detects minimal residual disease (MRD) in pediatric acute lymphoblastic leukemia (ALL). This rapid method aids in identifying relapse risk for T-lineage ALL patients with the TAL-1 deletion.
Area of Science:
- Hematology
- Molecular Biology
- Pediatric Oncology
Background:
- Hematologic relapse is a major challenge in curing childhood acute lymphoblastic leukemia (ALL).
- Minimal residual disease (MRD) detection during remission can identify patients at increased risk of relapse.
- Current polymerase chain reaction (PCR) methods for MRD assessment are complex and time-consuming.
Purpose of the Study:
- To develop an improved, simplified assay for MRD assessment in pediatric ALL.
- To establish a real-time quantitative PCR (RQ-PCR) assay for detecting leukemic cells with the TAL-1 deletion.
Main Methods:
- Development of a TAL-1 specific real-time quantitative PCR (RQ-PCR) assay.
- Validation of assay sensitivity using serial dilutions of leukemic DNA (down to 1 in 100,000 cells).
- Comparison of TAL-1 RQ-PCR with limiting dilution analysis on 23 samples from eight pediatric ALL patients in remission.
Main Results:
- The TAL-1 RQ-PCR assay demonstrated high sensitivity, detecting one leukemic cell among 100,000 normal cells.
- Both methods detected MRD levels ≥0.001% in 17 out of 23 samples, showing good correlation (r² = 0.926).
- Fewer than 0.001% leukemic cells were detected by both methods in the remaining six samples.
Conclusions:
- The TAL-1 RQ-PCR assay provides a rapid, sensitive, and accurate method for MRD assessment.
- This assay is suitable for T-lineage ALL patients with the TAL-1 deletion.
- Improved MRD assessment can aid in managing pediatric ALL and reducing relapse rates.
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