Abstract
Objective:To optimize and evaluate a real time PCR of Single Nucleotide Polymorphism by SYBR Green method for detection of donor chimerism after haematopoietic stem cell transplantation.Methods:This descriptive study was conducted at Genetic Resource Centre (GRC) Lab Rawalpindi from Oct 2017 - Dec 2019. A total of twenty patients of post haematopoietic stem cell transplant with various haematological disorders were studied to see the status of donor chimerism by using SNP real time PCR using SYBR Green method and short tandem repeat PCR. These patients had undergone allogeneic HSCT from HLA-matched sibling donors at Pakistan Institute of Medical Science and Armed Forces Bone Marrow Transplant Centre.Results:Real time PCR using SYBR Green was able to detect significant amount of chimerism in all 20 patients having undergone HSCT. Regarding precision of the real time PCR assay the mean value of donor chimerism was 94.1% (SD 3.96) and by STR PCR it was 95.1% (SD 1.41). The assay was found to be sensitive with a detection limit of <1%.Conclusion:Our results demonstrate that SNP analysis by SYBR Green real time PCR may be used for the evaluation of chimerism status in patients having undergone HSCT with a sensitivity of <1%. Hence donor chimerism by this sensitive method can be used in monitoring of chimerism in post-transplant patients with various haematological disorders.
Highlights
Haematopoietic stem cell transplantation (HSCT) has been extensively used for patients suffering from malignant hematological or nonmalignant hematological diseases,[1] and considered as a1
Results of donor chimerism by real time SNP PCR in the 20 sibling pairs were recorded in the form of PCR plots
Results of donor chimerism by STR PCR in the 20 sibling pairs were read from the silver-stained polyacrylamide gels and their densitometry
Summary
Haematopoietic stem cell transplantation (HSCT) has been extensively used for patients suffering from malignant hematological or nonmalignant hematological diseases,[1] and considered as a1. Post-transplant monitoring in HSCT is used to predict relapse status, graft rejection and GVHD, in order to tailor the appropriate therapy.[4]
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