Quantification of chimerism within peripheral blood, bone marrow and purified leukocyte subsets: comparison of singleplex and multiplex PCR amplification of short tandem repeat (STR) loci.
OBJECTIVE
Chimerism analysis has become a routine diagnostic procedure after haematopoietic allogeneic stem cell transplantation for early detection of relapse of disease or graft failure. Whereas some centres developed individual in-house short tandem repeat (STR) systems, others prefer commercial multiplex PCR systems. However, little is known about inter-assay variation, which could have a significant impact on treatment decision. We therefore compared two commercial multiplex PCR kits with our in-house STR system using different sample sources, such as peripheral blood (PB), bone marrow (BM) and specific leukocyte subsets.


RESULTS
Fifty samples of eighteen paediatric patients were analysed. For neither material, PB, BM and leukocyte subtypes, a significant difference between the STR systems tested was observed. Chimerism analyses of each single STR primer, which is component of both the in-house and the commercial STR system, did not reveal significant differences.


CONCLUSIONS
Our analysis demonstrates that similar results can be obtained with both assays, even when using various sample sources. Further evaluation of different test systems will help to increase interlaboratory standardisation of chimerism analyses for early clinical intervention.
Beck
O
O
Paediatric Haematology and Oncology, J.W. Goethe University Frankfurt/Main, Germany.
Seidl
C
C
Lehrnbecher
T
T
Kreyenberg
H
H
Schwabe
D
D
Klingebiel
T
T
Seifried
E
E
Bader
P
P
Koehl
U
U
D000293:Adolescent; D000328:Adult; D001769:Blood; D001853:Bone Marrow; D002648:Child; D002675:Child, Preschool; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007107:Immune System / Q000166:cytology; D007962:Leukocytes; D016131:Lymphocyte Subsets; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012038:Regeneration; D020080:Tandem Repeat Sequences; D018183:Transplantation Chimera; D014184:Transplantation, Homologous