Validation of chimerism in pediatric recipientsof allogeneic hematopoietic stem cell transplantation (HSCT) a comparison betweentwo methods: Real-time PCR (qPCR) vs. variable number tandem repeatsPCR (VNTR PCR)

摘要

Post-hematopoietic stem cell transplantation (HSCT) chimerism monitoring is important to assess relapse and therapeutic intervention. The purpose of our study is to compare two methods variable number tandem repeats (VNTR) vs. quantitative real-time polymerase chain reaction (qPCR) in terms of determining chimerism. 127 (peripheral blood n = 112, bone marrow n = 15) samples were simultaneously tested by VNTR using APO-B, D1S80, D1S111, D17S30, gene loci SRY and ZP3 and qPCR using 34 assays (CA001-CA034) that are designed to a bi-allelic insertion/deletion (indel) polymorphism in the human genome. Samples were separated in three subsets: total WBC, T-cell and myeloid cells. Extraction of DNA was performed then quantified. We analyzed column statistics, paired t-test and regression analysis for both methods. There was complete correlation between the two methods. The simplicity and rapidity of the test results from the qPCR method is more efficient and accurate to assess chimerism.