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Updated: Aug 8, 2026
VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
Published on: December 28, 2015
Immunoglobulin heavy-chain consensus probes for real-time PCR quantification of residual disease in acute
J W Donovan1, M Ladetto, G Zou
1Department of Adult Oncology, Harvard Medical School, Dana-Farber Cancer Institute, Boston, MA 02115, USa. john_william_donovan@dfci.harvard.edu
Insights
Detecting minimal residual disease (MRD) in B-cell malignancies using immunoglobulin heavy-chain (IgH) rearrangements is crucial. A new real-time PCR method accurately quantifies MRD, identifying relapse risk in acute lymphoblastic leukemia (ALL) patients.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- Immunoglobulin heavy-chain (IgH) rearrangements serve as key markers for detecting minimal residual disease (MRD) in B-cell malignancies via polymerase chain reaction (PCR).
- Qualitative PCR detection of MRD in childhood acute lymphoblastic leukemia (ALL) has limitations in predicting relapse, as prolonged persistence of detectable disease does not always correlate with adverse outcomes.
- Quantitative assessment of tumor burden is increasingly recognized as a critical factor for identifying high-risk ALL patients.
Purpose of the Study:
- To develop an efficient and reliable real-time PCR method for quantifying MRD in ALL patients.
- To establish a sensitive assay capable of discriminating between patients with persistent MRD who relapse and those who do not.
Main Methods:
- Development of a real-time PCR assay utilizing immunoglobulin heavy-chain (IgH) V(H) gene family consensus fluorogenically labeled probes for MRD quantification.
- Validation of the assay's accuracy and reproducibility across a wide range of tumor burdens.
- Assessment of the method's sensitivity, capable of detecting approximately 1 tumor cell in 5 x 10(4) normal cells.
Main Results:
- The developed real-time PCR method accurately and reproducibly quantifies MRD.
- The assay demonstrates high sensitivity, detecting minimal residual disease at a level of 1 tumor cell in 5 x 10(4) normal cells.
- This quantitative approach successfully differentiates between ALL patients with persistent MRD who relapse and those who remain relapse-free.
Conclusions:
- The novel real-time PCR technique provides an efficient and reliable means for quantifying MRD in ALL patients.
- This methodology is broadly applicable to various B-cell malignancies for MRD monitoring.
- The ability to quantify MRD improves risk stratification and has significant implications for clinical management and prospective studies.
Abstract:
Tumor-related immunoglobulin heavy-chain (IgH) rearrangements are markers for polymerase chain reaction (PCR) detection of minimal residual disease (MRD) in B-cell malignancies. Nested PCR with patient IgH allele-specific oligonucleotide primers can detect 1 tumor cell in 10(4) to 10(6) normal cells. In childhood acute lymphoblastic leukemia (ALL), persistence of PCR-detectable disease is associated with increased risk of relapse. The clinical significance of qualitative PCR data can be limited, however, because patients can harbor detectable MRD for prolonged periods without relapse. Recent studies indicate that a quantitative rise in tumor burden identifies patients who are at high risk for relapse. Therefore, an efficient and reliable PCR method for MRD quantification is needed for ALL patients. We have developed a real-time PCR method to quantify MRD with IgH V(H) gene family consensus fluorogenically labeled probes. With this method, a small number of probes can be used to quantify MRD in a large number of different patients. The assay was found to be both accurate and reproducible over a wide range and capable of detecting approximately 1 tumor cell in 5 x 10(4) normal cells. We demonstrate that this methodology can discriminate between patients with persistence of MRD who relapse and those who do not. This technique is generally applicable to B-cell malignancies and is currently being used to quantify MRD in a number of prospective clinical studies at our institution. (Blood. 2000;95:2651-2658)
