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Blood, 1 April 2001, Vol. 97, No. 7, pp. 1929-1936
CLINICAL OBSERVATIONS, INTERVENTIONS, AND THERAPEUTIC TRIALS
Clonotypic polymerase chain reaction confirms minimal residual
disease in CLL nodular PR: results from a sequential treatment
CLL protocol
Ariela Noy,
Ravi Verma,
Martha Glenn,
Peter Maslak,
Zia U. Rahman,
James R. Keenan,
Mark Weiss,
Daniel Filippa, and
Andrew D. Zelenetz
From the Department of Medicine/Division of
Hematologic Oncology and the Department of Pathology, Memorial
Sloan-Kettering Cancer Center, New York, NY.
Patient-tumor-specific oligonucleotides were generated for the
detection of minimal residual disease (MRD) in a highly specific and
sensitive clonotypic polymerase chain reaction (cPCR). The clone-specific region of highest diversity, CDR-III, was PCR amplified and sequenced. Nested CDR-III clonotypic primers were used in a
semi-nested cPCR with a sensitivity of at least 1 in 105
cells. Patients with protocol-eligible Rai intermediate or high-risk chronic lymphocytic leukemia (CLL) received induction with fludarabine 25 mg/m2 per day for 5 days every 4 weeks for 6 cycles,
followed by consolidative high-dose cyclophosphamide (1.5, 2.25, or
3g/m2). cPCR was performed on peripheral blood and bone
marrow mononuclear cells. All 5 patients achieving a clinical partial
remission (PR) studied by cPCR were positive. Five patients
achieved nodular PR (nPR) (residual nodules or suspicious
lymphocytic infiltrates in a bone marrow biopsy as the sole suggestion
of residual disease). Five of 5 patients with nPR were cPCR positive.
In contrast, flow cytometry for CD5-CD19 dual staining and  -
clonal excess detected MRD in only 3 of the same 5 nPR patients, all of
whom were cPCR positive, and immunohistochemistry detected MRD in only
1 of 4 assessable patients. Three of 7 CR patients evaluable by cPCR had MRD. Only 1 CR patient had MRD by flow cytometry; that patient was
also cPCR positive. These data support the conclusions that nodular PR
in CLL represents MRD and that clonotypic PCR detects MRD in CLL more
frequently than flow cytometry or immunohistochemistry.
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