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Detection of Microchimerism After Allogeneic Blood
Transfusion Using Nested Polymerase Chain Reaction Amplification With
Sequence-Specific Primers (PCR-SSP): A Cautionary Tale
Anthony S. Carter,
Mike Bunce,
Lucia Cerundolo,
Ken I. Welsh,
Peter J. Morris, and
Susan V. Fuggle
From the Nuffield Department of Surgery, University of Oxford, John
Radcliffe Hospital, Headington, Oxford, UK.
In bone marrow transplantation, the detection of chimerism is an
important adjunct to the repertoire of tests available for determining
acceptance of the graft. In solid organ transplantation, there is
currently intense interest in the role that chimerism plays in both
short- and long-term host reactivity to the graft. Allogeneic blood
transfusion has been associated with a subtle immunosuppressive effect
in renal transplantation and chimerism is implicated as a possible
mechanism for this effect. To assess the survival of allogeneic cells
after blood transfusion or transplantation, we have developed a
technique based on molecular typing for HLA class II alleles, which
enables the detection of donor-derived cells in patients receiving
blood transfusions. While developing this technology, we investigated
why we and others observe false amplification. Sequencing of false
products has shown that they arise from amplification of both
pseudogenes and non-pseudogenes present in the DNA under test.
Elucidation of this phenomenon allows the amplification of these false
products to be predicted in any given combination and hence avoided by
the judicious selection of primers. Validation has been achieved by
following donor alleles after transfusion of blood containing defined
numbers of leukocytes expressing selected mismatched antigens.
Blood, Vol. 92 No. 2 (July 15), 1998:
pp. 683-689
© 1998 by the American Society of Hematology.
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