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Chimerism Monitoring after Allogeneic Hematopoietic Stem Cell Transplantation Using Quantitative Real-Time PCR of Biallelic Insertion/Deletion Polymorphisms Seon Young Kim, Moon Hwan Jeong, Nare Park, Eunkyoung Ra, Hyunwoong Park, Soo Hyun Seo, Ji Yeon Kim, Moon-Woo Seong, Sung Sup Park The Journal of Molecular Diagnostics Volume 16, Issue 6, Pages (November 2014) DOI: /j.jmoldx Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions
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Figure 1 AlleleSEQR Chimerism Assay results for markers CA014, CA027, and CA030 using 250 ng (A and B) and 25 ng (C and D) of input DNA. These plots specifically show lower ranges of recipient DNA with 250 ng (B) and 25 ng (D) input DNA. The black diagonal lines represent summarized regression analysis results of all three markers. The Journal of Molecular Diagnostics , DOI: ( /j.jmoldx ) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions
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Figure 2 Comparison of recipient DNA percentages obtained between the AlleleSEQR Chimerism Assay and the STR (A and B) and the X/Y fluorescence in situ hybridization (FISH) method (C and D) in 175 samples after transplantation. Gray circles represent recipient DNA percentages; black lines, regression; and dashed lines, 95% CI of regression analysis. The Journal of Molecular Diagnostics , DOI: ( /j.jmoldx ) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions
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Figure 3 The recipient DNA percentages as measured by the AlleleSEQR Chimerism Assay and the STR method in acute leukemia patients who relapsed during the follow-up period (A) and those who did not relapse (B). Black arrows indicate when the relapse of acute leukemia was diagnosed; white triangles, the timing of bone marrow biopsy studies performed in remission status; black triangles, when bone marrow studies showed persistence of leukemic cells. The Journal of Molecular Diagnostics , DOI: ( /j.jmoldx ) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions
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Figure 4 Variation in CT values of reference and test samples among different specimens. In 14 samples in which two quantitative AlleleSEQR Chimerism Assays were performed, the difference in CT value between the first and second test was plotted for each patient. The CT values of the specific marker or positive control (CA999) show wide variations and moves in similar directions. The calculated ΔCT of the reference sample (CT informative marker of pre-HSCT sample − CT CA999 of pre-HSCT sample) has lower variability. The Journal of Molecular Diagnostics , DOI: ( /j.jmoldx ) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions
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Figure 5 Estimation of quantitative AlleleSEQR Chimerism Assay results using 1.03 as the estimated ΔCT value for the reference samples. A: Quantitative AlleleSEQR Chimerism Assay results, estimated AlleleSEQR Chimerism Assay results, and STR results for total samples. B: Results for samples with recipient DNA percentages ranging from 0% to 2%. The Journal of Molecular Diagnostics , DOI: ( /j.jmoldx ) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions
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