Abstract
BackgroundAllogeneic hematopoietic stem cell transplantation (allo-HSCT) is performed mainly in patients with high-risk or advanced hematologic malignancies and congenital or acquired aplastic anemias. In the context of the significant risk of graft failure after allo-HSCT from alternative donors and the risk of relapse in recipients transplanted for malignancy, the precise monitoring of posttransplant hematopoietic chimerism is of utmost interest. Useful molecular methods for chimerism quantification after allogeneic transplantation, aimed at distinguishing precisely between donor's and recipient's cells, are PCR-based analyses of polymorphic DNA markers. Such analyses can be performed regardless of donor's and recipient's sex. Additionally, in patients after sex-mismatched allo-HSCT, fluorescent in situ hybridization (FISH) can be applied.MethodsWe compared different techniques for analysis of posttransplant chimerism, namely FISH and PCR-based molecular methods with automated detection of fluorescent products in an ALFExpress DNA Sequencer (Pharmacia) or ABI 310 Genetic Analyzer (PE). We used Spearman correlation test.ResultsWe have found high correlation between results obtained from the PCR/ALF Express and PCR/ABI 310 Genetic Analyzer. Lower, but still positive correlations were found between results of FISH technique and results obtained using automated DNA sizing technology.ConclusionsAll the methods applied enable a rapid and accurate detection of post-HSCT chimerism.
Highlights
Allogeneic hematopoietic stem cell transplantation is performed mainly in patients with high-risk or advanced hematologic malignancies and congenital or acquired aplastic anemias
5, 6, 7) only donor's cells were detected in all post-HSCT samples and in one patient only recipient's cells were present in all samples examined
Three other patients exhibited mixed posttransplant chimerism according to PCR and/or fluorescent in situ hybridization (FISH)
Summary
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is performed mainly in patients with high-risk or advanced hematologic malignancies and congenital or acquired aplastic anemias. Useful molecular methods for chimerism quantification after allogeneic transplantation, aimed at distinguishing precisely between donor's and recipient's cells, are PCR-based analyses of polymorphic DNA markers Such analyses can be performed regardless of donor's and recipient's sex. Accepted molecular methods for analysis of chimerism after allo-HSCT, aimed at distinguishing precisely between donor's and recipient's cells, are PCR-based analyses of polymorphic DNA markers, such as variable number of tandem repeats (VNTR) or short tandem repeats (STR) [1]. Such analyses can be performed regardless of donor's and recipient's sex [2]. Successful outcome has been associated with a state of stable complete chimerism [4]
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