Purpose: To develop a standardized real-time polymerase chain reaction (PCR) method of quantifying minimal residual disease (MRD) in patients with pre-B acute lymphoblastic leukemia (ALL). Patients and Methods: In a series of 24 follow-up bone marrow (BM) samples in 11 patients (14 clonal markers), we performed real-time PCR assays using one consensus and one clone-specific primer for each marker. The markers analyzed included immunoglobulin heavy chain (IgH), T-cell receptor (TCR) and TEL-AML1 rearrangements. Results: We achieved a detection limit of 3.3 × 10-5 ± 1.2 × 10-5 and an accurate quantitation (r = -0.99) limit of 2.0 × 10-4 ± 8.8 × 10-5 blasts. Both inter- and intra-assay reproducibility were exceptional. Additionally, we found comparable results to those of a "gold standard" limiting-dilution PCR assay (r = 0.62). Conclusions: The IgH, TCR and TEL-AML1 markers can be used as targets by real-time PCR under the same cycling profile, allowing quantitation of MRD in more 95% of patients with pre-B ALL. This standardized, real-time PCR technique should simplify monitoring MRD in clinical trials.
All Science Journal Classification (ASJC) codes
- Pediatrics, Perinatology, and Child Health
- Minimal residual disease
- Pediatric acute lymphoblastic leukemia
- Real-time quantitative PCR