کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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5666342 | 1591375 | 2017 | 11 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: It's about time: The development and validation of a rapid optimized single antigen bead (ROB) assay protocol for LABScreen It's about time: The development and validation of a rapid optimized single antigen bead (ROB) assay protocol for LABScreen](/preview/png/5666342.png)
The LABScreen single antigen bead assay (SAB) is a method widely used for the identification and monitoring of human leukocyte antigen (HLA) antibodies in patients pre-and post-transplant. While accurate testing of patient samples is key for optimal patient care, time can also be important, especially during deceased donor workups or post-transplant assessments. Here we describe the development and validation of the Rapid Optimized SAB (ROB) protocol, a modified version of the One Lambda LABScreen SAB (OLSAB) procedure, which reduces assay time from 85 to 25Â min (>70% reduction) without impacting assay quality or sensitivity. Optimization steps included shortened centrifugation cycles and reduced serum and secondary antibody incubation times in combination with increased secondary antibody concentration. Linear regression analysis of baseline median fluorescence intensity (MFI) values showed excellent correlation between the ROB and OLSAB protocols (r2Â >Â 0.98) for both class I and class II antibodies in 58 sera tested in two HLA laboratories. Importantly, the ROB protocol demonstrated a trend towards improved inter-laboratory MFI concordance when compared to the OLSAB procedure (r2Â =Â 0.9816 vs 0.9451), especially for HLA antibody specificities in the 500-2000 MFI range (r2Â =Â 0.7824 vs 0.6313). Implementation of the ROB protocol will expedite HLA antibody testing and may improve reproducibility of the SAB assay.
Journal: Human Immunology - Volume 78, Issues 7â8, JulyâAugust 2017, Pages 489-499