Analysis of hematopoietic chimerism in patients suffering from leukaemia after allogeneic stem cell transplantation
biallelic polymorphism ; STR ; STR-PCR ; RQ-PCR ; hematopoietic chimerism
Allogeneic hematopoietic stem cell transplantation (alloSCT) is a process that requires the engraftment of transplanted hematopoietic stem cells which re-establish normal hematological and immunological systems. One of the most useful tool for engraftment monitoring is the assessment of hematopoietic chimerism. which occurs after alloSCT and describes the percentage of donor hematopoietic and lymphoid cells in a transplant recipient. 38 patients after alloSCT entered the study and the total number of transplantations was 43. Hematopoietic chimerism evaluations were performed on 30, 60, 90, 120, 150, 180, 270 and 360 day post alloSCT. The evaluation of hematopoietic chimerism was based on PCR amplification of polymorphic non-coding DNA sequences - short tandem repeats (STR-PCR). The main tool was a semi-quantitive method – fragment length analysis. The product of amplification was analyzed using the sequencer. The second method was based on a quantitative Real Time PCR technique (RQ-PCR). There were performed amplification of biallelic non-coding DNA sequences with short insertions or deletions. STR-PCR and RQ-PCR chimerism assays were compared and results evidenced the greater sensitivity of RQ-PCR method. There were not crucial differences in the results of chimerism evaluation obtained by means of these two methods. The analysis of chimerism kinetics allowed to modify the pos ; t-transplantation-treatment in 3 patients: 2 decision of DLI, 1 withdrawal of CsA, 1 pt giving G-CSF. The results of chimerism monitoring confirmed that the failure of achieving a CC or lost of CC can predict the relapse of the disease.
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hematologia ; transplantologia