Detection of Microchimerism by Minor Histocompatibility Antigen HA-1 Allele-Specific Nested Polymerase Chain Reaction Brigitte Wieles, Jos Pool, Richard.

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Detection of Microchimerism by Minor Histocompatibility Antigen HA-1 Allele-Specific Nested Polymerase Chain Reaction Brigitte Wieles, Jos Pool, Richard Derks, William J. Burlingham, Els Goulmy Biology of Blood and Marrow Transplantation Volume 11, Issue 5, Pages 345-348 (May 2005) DOI: 10.1016/j.bbmt.2005.01.009 Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Scheme of part of the HA-1-encoding locus, indicating primer positions used for the development of the HA-1 allele-specific nested PCR. Biology of Blood and Marrow Transplantation 2005 11, 345-348DOI: (10.1016/j.bbmt.2005.01.009) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Sensitivity determination of the HA-1 nested PCR. A, FIIH as the forward primer in the second PCR step. B, FIIR as the forward primer in the second PCR step. M indicates marker; lane 1, no template control; lanes 2 to 9, PCR results of serial dilutions of cells homozygous for HA-1H diluted in cells homozygous for HA-1R in a ratio of 1:10 to 1:108; lane 10, homozygous HA-1H control sample; lane 11, homozygous HA-1R control sample. Biology of Blood and Marrow Transplantation 2005 11, 345-348DOI: (10.1016/j.bbmt.2005.01.009) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Analysis of a clinical sample for the presence of HA-1 microchimerism. Nested PCR results of a sample derived from an HA-1R kidney transplantation patient receiving an HA-1H kidney graft. A, FIIH as the forward primer in the second PCR step. B, Positive control PCR using FIIR as the forward primer in the second PCR step. Lanes 1 to 8 indicate PCR results of serial dilution of HA-1H cells in HA-1R cells in a ratio of 1:10 to 1:108; lane 9, homozygous H control sample; lane 10, homozygous R control sample; lane 11, T-cell fraction; lane 12, DC1 cell fraction; lane 13, DC2 cell fraction; lane 14, B-cell fraction; lanes 15 and 16, no-template control of the first PCR; lane 17, no-template control of the second PCR; M, marker. Biology of Blood and Marrow Transplantation 2005 11, 345-348DOI: (10.1016/j.bbmt.2005.01.009) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions