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- W234750149 abstract "The LABScreen single antigen bead (SAB) assay is routinely used for HLA antibody detection and allows for testing of multiple patient samples. Despite this, standard SAB assay is time intensive, taking 1.5–2 hr to perform, which is not optimal for urgent testing. Utilizing a multicenter study we compared the rapid optimized SAB (ROB) protocol (Liwski et al. ASHI 2014), with a 30 min turn-around time (TAT), to standard SAB LABScreen method. Ten HLA labs (BCM, DCI, Edmonton, Halifax, Georgetown, Winnipeg, METC, PSBC, SWI, UPMC) participated in the study. Five ASHI 2014 AC-1 survey sera (AC 460-464) were tested in each lab with standard SAB LABScreen (SSL) vs the ROB protocol. Class I LS1A04 (lot 8) and class II LS2A01 (lot 10) HLA kits were used. Mean fluorescence intensity (MFI) values were collected. Protocols were compared with respect to the mean MFI, specificities reaching consensus (⩾90% labs; 2000 MFI cutoff) and coefficient of variation (%CV). HLA antibody reactivity patterns were similar between the protocols with an excellent MFI correlation (r2>0.99) and signal to noise differential. Average MFI was slightly reduced with the ROB protocol (by 5.0%). The number of specificities reaching consensus (⩾90% labs; 2000 MFI cutoff) was similar between protocols (97 SSL vs 94 ROB). When the ROB protocol cutoff was lowered to 1500 MFI, the number of consensus specificities improved to 99, including all 97 SSL specificities (Fig. 1). Importantly, the average %CV of raw bead MFI was significantly improved with the ROB protocol (ROB: 21.2+/−3.4 vs SSL: 34.4+/−5.6; p< 0.001; Fig. 2). A slight increase in background was noted with the ROB protocol (AC 462 serum) in some labs. However, this had no impact on antibody assignment and improved with one additional wash step. ROB protocol reduces SAB LABScreen assay time by 70% and improves precision without compromising sensitivity or specificity. Implementation of the ROB protocol will expedite antibody testing in urgent cases and improve TAT for routine testing." @default.
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- W234750149 date "2015-04-01" @default.
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- W234750149 title "LBP11" @default.
- W234750149 doi "https://doi.org/10.1016/j.humimm.2015.01.043" @default.
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