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Lot‐to‐lot variability in HLA antibody screening using a multiplexed bead‐based assay
Author(s) -
Gandhi Manish J.,
Carrick Danielle M.,
Jenkins Sarah,
De Goey Steven,
Ploeger Nancy A.,
Wilson Gregory A.,
Lee Jar How,
Winters Jeffrey L.,
Stubbs James R.,
Toy Pearl,
Norris Philip J.
Publication year - 2013
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.12064
Subject(s) - confidence interval , multiplex , antibody , medicine , human leukocyte antigen , antigen , immunology , biology , genetics
Background Identifying antibodies to HLA (anti‐ HLA ) by solid‐phase assays is used to screen blood donors to mitigate transfusion‐related acute lung injury risk. Various cutoffs for detection assays have been proposed in the literature; however, these do not take into consideration lot‐to‐lot variability of commercially available assays. Study Design and Methods Samples from 93 nontransfused males were tested using five different lots of a multiplex bead‐based anti‐ HLA detection kit. A subset of 17 samples was tested on 5 days using a single lot. An additional 96 samples from donations with varied anti‐ HLA levels were tested using kits from two different lots. Results were reported as a normalized background ( NBG ) ratio. Results For the 93 nontransfused donors, NBG values generated using the reference lot were significantly higher than those obtained with three of the four comparator lots. However, for the 96 samples with low‐, moderate‐, and higher‐level anti‐ HLA , Class I (CL‐I) values were 1.4 times lower and Class II (CL‐II) values were 1.2 times lower using the reference versus comparator lot. For CL‐I antibodies the between‐lot standard deviation ( SD ) was 1.36 (95% confidence interval [ CI ], 1.19‐1.60), while the between‐day SD was 1.27 (95% CI , 1.08‐1.52). Similarly, for CL‐II antibodies the between‐lot SD was 0.81 (95% CI , 0.70‐0.95), while the between‐day SD was 0.50 (95% CI , 0.43‐0.60). Conclusions There is interlot variability in the tested HLA detection assay as well as significant bias between lots. It may be reasonable to develop a new cutoff when a new lot is obtained.