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Transplantation. 2019 Aug 1. doi: 10.1097/TP.0000000000002900. [Epub ahead of print]

DSA-FXM: Accelerated donor specific flow crossmatch discriminating class I and II antibody specifically and only to donor HLA for determining true incompatibility.

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Histocompatibility, Immunogenetics & Disease Profiling Laboratory, Stanford Blood Center, 3373 Hillview Avenue, Palo Alto, CA, USA 94304.
Department of Pathology, Stanford University School of Medicine, 3373 Hillview Avenue, Palo Alto, CA, USA 94304.



Worldwide, a final crossmatch is the gold standard for determining compatibility between patient and donor prior to solid organ transplantation and preventing hyperacute rejection. In the absence of autoantibodies, an incompatible crossmatch in a sensitized patient is attributed to mismatched donor human leukocyte antigens (HLA). However, current physical crossmatch methods cannot distinguish reactivity to HLA from other clinically irrelevant cell surface targets nor the class of HLA if it is the target. Result interpretation is difficult or impossible when autoantibodies, alloantibodies, and/or therapeutic antibodies coexist.


Herein we describe a unique donor specific flow crossmatch (DSA-FXM) that distinguishes HLA class I and/or II DSA bound to HLA antigens on the donor cell surface in their native conformation that is not impacted by rituximab, ATG (after absorption), or auto-antibodies. It is HLA specific.


We compared results of single antigen antibody testing, auto- and allo-FXM using traditional FXM and our DSA-FXM method from 94 patients (enriched for auto+/allo+ pairs; n=64) against 110 donors (338 tests) and show that, in our cohort, positive traditional FXM results are not directed to donor HLA 60.25% of the time, and negative traditional FXM results are missing HLA DSA 36.2% of the time based on the DSA-FXM.


We demonstrate that the DSA-FXM is able to define categorically distinct and clinically important HLA antibody profiles in half the time required for the standard FXM, potentially shortening cold ischemia time and providing clinicians with unambiguous essential information regarding HLA compatibility when time is critical.

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