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Blood, Vol. 96 No. 1 (July 1), 2000:
pp. 63-70
Consensus strategy to quantitate malignant cells in myeloma
patients is validated in a multicenter study
Peter Willems,
Onno Verhagen,
Christine Segeren,
Peter Veenhuizen,
Jeroen Guikema,
Erik Wiemer,
Laura Groothuis,
Tineke
Buitenweg-de Jong,
Henriëtte Kok,
Andries Bloem,
Nico Bos,
Edo Vellenga,
Ewald Mensink,
Pieter Sonneveld,
Henk Lokhorst Ellen van der Schoot, and
Reinier Raymakers; for the
Belgium-Dutch Hematology-Oncology Group
From the Department of Hematology, Academic Hospital
Rotterdam; the Department of Hematology, Erasmus University Rotterdam;
the Department of Hematology, Academic Hospital Nijmegen; the
Departments of Hematology and Immunology, Academic Hospital Utrecht;
the Department of Hematology, Academic Hospital Groningen; the
Department of Immunohematology, Central Laboratory of the Netherlands
Red Cross Blood Transfusion Service; and the Department of Histology
and Cell Biology, University of Groningen, the Netherlands.
Recently the Belgium-Dutch Hematology-Oncology group initiated a
multicenter study to evaluate whether myeloma patients treated with
intensive chemotherapy benefit from additional peripheral stem cell
transplantation. To determine treatment response accurately, we decided
to quantitate malignant cells. To test a consensus quantitation
strategy, 5 centers independently determined the immunoglobulin heavy
chain sequences of patient tumor cells and developed allele-specific
oligonucleotides (ASO) and ASO-polymerase chain reaction (PCR). We
compared the reproducibility of real-time quantitation with
quantitation using limiting dilutions. We distributed DNA samples with
a 4-log range of tumor cell concentrations and found average
quantitation values deviating 74% and 42% from the input values with
real-time PCR (1 center) and limiting dilutions (4 centers),
respectively. Within single centers we found an average variation
coefficient of 0.74, with limiting dilutions not significantly different from the average 0.82 center-to-center variation coefficient. Within a single center, real-time quantitation proved more reproducible (average variation coefficient, 0.36). Quantification was confirmed in
3 patients during treatment in the protocol. This report shows that
real-time PCR or limiting dilution assays can be used for quantitation
in a single multicenter trial. We present a consensus strategy that
allows an accurate comparison of quantitation data generated in
independent centers.
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