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Analysis of B-lymphoid malignancies using allele-specific polymerase chain
reaction: a technique for sequential quantitation of residual disease
D Billadeau, M Blackstadt, P Greipp, RA Kyle, MM Oken, N Kay and B Van Ness
Department of Biochemistry, University of Minnesota, Minneapolis 55455.
The junctional sequences corresponding to the complementarity determining
region 3 (CDR3) of rearranged heavy chain Ig genes can provide
allele-specific markers in the detection of B-lymphoid malignancies.
Consensus oligonucleotide primers were used to amplify CDR3 regions of
rearranged heavy chain alleles in clinical samples from myeloma, acute
lymphocytic leukemia, and chronic lymphocytic leukemia patients. From the
sequence of the amplified products, allele-specific primers were
synthesized and used directly in polymerase chain reaction (PCR)
amplification to detect the malignant clone. This method was both highly
specific and sensitive to 1 malignant B-cell in a background of 10(5)
normal cells. In addition, parameters that affect the linearity of PCR
detection were determined and, by using titrations of malignant target
cells to generate standard curves, quantitations of residual malignancies
were determined. The application of this method is shown in an analysis of
myeloma patients whose marrows were analyzed sequentially during therapy.
Allele-specific oligonucleotide-PCR provided a rapid, highly specific and
quantitative measure of residual disease, even in patients with clinical
parameters indicating complete remission.
Volume 78,
Issue 11,
pp. 3021-3029,
12/01/1991
Copyright © 1991 by The American Society of Hematology
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