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Prepublished online as a Blood First Edition Paper on August 29, 2002; DOI 10.1182/blood-2002-05-1365.
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Blood, 1 January 2003, Vol. 101, No. 1, pp. 363-369
TRANSPLANTATION
A novel rapid single nucleotide polymorphism (SNP)-based method
for assessment of hematopoietic chimerism after allogeneic stem cell
transplantation
Ephraim P. Hochberg,
David
B. Miklos,
Donna Neuberg,
Daniel A. Eichner,
Stephen F. McLaughlin,
Alex Mattes-Ritz,
Edwin P. Alyea,
Joseph H. Antin,
Robert J. Soiffer, and
Jerome Ritz
From the Center for Hematologic Oncology and the
Department of Biostatistics, Dana-Farber Cancer Institute,
Department of Medicine, Brigham & Women's Hospital, Harvard Medical
School, Boston, MA.
A major end point of nonmyeloablative hematopoietic stem
cell transplantation is the attainment of either mixed
chimerism or full donor hematopoiesis. Because the
majority of human genetic disparity is generated by single nucleotide
polymorphisms (SNPs), direct measurement of SNPs should provide a
robust tool for the detection and quantitation of chimerism. Using
pyrosequencing, a rapid quantitative sequencing technology, we
developed a SNP-based assay for hematopoietic chimerism. Based on 14 SNPs with high allele frequencies, we were able to identify at
least 1 informative SNP locus in 55 patients with HLA-identical donors.
The median number of informative SNPs in related pairs was 5 and in
unrelated pairs was 8 (P < .0001). Assessment of
hematopoietic chimerism in posttransplantation DNA was shown to be
quantitative, accurate, and highly reproducible. The presence of
5% donor cells was reliably detected in replicate assays.
Compared with current measures of engraftment based on identification
of short tandem repeats (STRs), variable number of tandem repeats
(VNTRs), or microsatellite polymorphisms, this
SNP-based method provides a more rapid and quantitative assessment of
chimerism. A large panel of SNPs enhances the ability to identify an
informative marker in almost all patient/donor pairs and also facilitates the simultaneous use of multiple markers to improve the
statistical validity of chimerism measurements. The inclusion of SNPs
that encode minor histocompatibility antigens or other genetic
polymorphisms that may influence graft-versus-host disease or other
transplantation outcomes can provide additional clinically relevant
data. SNP-based assessment of chimerism is a promising technique that
will assist in the analysis of outcomes following transplantation.
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