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Blood, Vol. 95 No. 8 (April 15), 2000:
pp. 2651-2658
Immunoglobulin heavy-chain consensus probes for real-time PCR
quantification of residual disease in acute lymphoblastic leukemia
John W. Donovan,
Marco Ladetto,
Guangyong Zou,
Donna Neuberg,
Christina Poor,
Dorothy Bowers, and
John G. Gribben
From the Department of Adult Oncology, Department of Medicine,
Harvard Medical School, and Department of Biostatistics, Dana-Farber
Cancer Institute, Boston, MA.
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
104 to 106 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
VH 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 × 104 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.
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