Universität zu Köln

Tumer, Gizem, Gniadek, Thomas, Baye, Jennifer, Pena, Ryan, Warner, Paul, Fung, Mark, Beaudin, Lynette, Dunckley, Heather, Gandhi, Manish ORCID: 0000-0001-7210-1909, Gathof, Birgit, Hsu, Susan, Klohe, Ellen, Marcus, Nalaja, Bamert, Roberta, Sims, Shona, Takanashi, Minoko, Wendel, Silvano and Cohn, Claudia S. (2020). The effect of serum pretreatment regimens for the detection of HLA class I antibodies in platelet-refractory patients. Transfusion, 60 (3). S. 488 - 498. HOBOKEN: WILEY. ISSN 1537-2995

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Abstract

BACKGROUND Single antigen bead (SAB) assays are used to identify human leukocyte antigen (HLA) antibodies in patients with platelet refractoriness due to HLA Class I alloimmunization. Some laboratories use serum pretreatment regimens to eliminate interference from immunoglobulin M antibodies and complement. These modifications may contribute to interlaboratory variability, which is a recognized problem with the SAB assay. STUDY DESIGN AND METHODS Five patients & apos; sera were overnight shipped to 12 laboratories in the United States and internationally. Recipients used their lab & apos;s SAB procedure to identify HLA Class I antibodies. The resultant mean fluorescence intensity (MFI) data were compared by instrumentation, bead lot, and pretreatment regimens. Laboratory-specific cutoffs for positive antibodies were applied to the results. RESULTS Interlaboratory variability for MFI values appears to be associated with different pretreatment regimens. The coefficient of variation (CV) of MFI from samples pretreated with ethylenediaminetetraacetic acid, dithiothreitol, or heat inactivation (EDHI) were similar, ranging from 14% to 56% (mean, 22%). For samples with no pretreatment, the CVs were significantly higher than EDHI-treated samples, ranging from 25% to 74% (mean, 39%; 95% confidence interval, 12.10-21.90; p < 0.0001). An intralaboratory comparison of pretreatment regimens confirmed these findings. Some positive antibody specificities present in EDHI-treated samples were negative in corresponding samples with no pretreatment when laboratory-specific cutoffs for positive antibodies were applied. CONCLUSION Our results show that greater interlaboratory precision can be achieved when samples are pretreated with EDHI as opposed to no pretreatment, likely because these pretreatments eliminate interference from inhibitors. Inhibitors may mask antibodies, leading to missed (or uncalled) specificities when no pretreatment is used.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Tumer, Gizem UNSPECIFIED UNSPECIFIED UNSPECIFIED Gniadek, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Baye, Jennifer UNSPECIFIED UNSPECIFIED UNSPECIFIED Pena, Ryan UNSPECIFIED UNSPECIFIED UNSPECIFIED Warner, Paul UNSPECIFIED UNSPECIFIED UNSPECIFIED Fung, Mark UNSPECIFIED UNSPECIFIED UNSPECIFIED Beaudin, Lynette UNSPECIFIED UNSPECIFIED UNSPECIFIED Dunckley, Heather UNSPECIFIED UNSPECIFIED UNSPECIFIED Gandhi, Manish UNSPECIFIED orcid.org/0000-0001-7210-1909 UNSPECIFIED Gathof, Birgit UNSPECIFIED UNSPECIFIED UNSPECIFIED Hsu, Susan UNSPECIFIED UNSPECIFIED UNSPECIFIED Klohe, Ellen UNSPECIFIED UNSPECIFIED UNSPECIFIED Marcus, Nalaja UNSPECIFIED UNSPECIFIED UNSPECIFIED Bamert, Roberta UNSPECIFIED UNSPECIFIED UNSPECIFIED Sims, Shona UNSPECIFIED UNSPECIFIED UNSPECIFIED Takanashi, Minoko UNSPECIFIED UNSPECIFIED UNSPECIFIED Wendel, Silvano UNSPECIFIED UNSPECIFIED UNSPECIFIED Cohn, Claudia S. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-348664
DOI:	10.1111/trf.15666
Journal or Publication Title:	Transfusion
Volume:	60
Number:	3
Page Range:	S. 488 - 498
Date:	2020
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1537-2995
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language INTERFERENCE; ASSAYS Multiple languages Hematology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/34866