Abstract：
Spherotech (SPT) microparticles capture non-specific binding materials present in test serum, and EDTA removes the so called\" prozone effect\". This study presents a novel approach of combined SPT-EDTA serum treatment prior to Luminex HLA antibody testing to remove high background, and prozone effect in a single step process, and compared the efficacy of SPT-EDTA serum pre-treatment with AdsorbOut (ADS) and Serum Cleaner (SC) to reduce background in solid phase immunoassays (SPI). A total of 21 serum samples with a history of elevated negative control (NC) values ≥500, and 20 samples with normal NC values were included to assess the potential adverse effects. A problem of high background was noted in 25% of our samples in SPI. We observed 80% effectiveness in reducing NC values <500 with SPT-EDTA serum pre-treatment compared to 72%, and 67% for ADS and SC-treated sera, respectively. Interestingly, we found a strong correlation in antibody-binding levels between SPT versus ADS; and ADS versus SC-treated sera for both phenotype and single antigen bead assays (p < 0.001). No adverse effect was noted on NC, positive control (PC) values, PC/NC ratios in the upfront use of SPT-EDTA as compared to EDTA alone. Our data revealed that combined SPT-EDTA treated sera is more effective than ADS, and SC in reducing high background in SPI. Taken together, SPT-EDTA serum treatment prior to Luminex HLA Ab testing is cost-effective, our laboratory saves nearly 30% of the annual total cost for Ab testing and improved test turnaround time by two business days.
摘要：
Spherotech（SPT）微粒捕获测试血清中存在的非特异性结合材料，EDTA消除了所谓的“前区效应”。这项研究提出了一种在LuminexHLA抗体测试之前联合SPT-EDTA血清治疗以去除高背景的新方法，和前区效应在一步过程中，并比较了用AdsorbOut（ADS）和血清清洁剂（SC）进行SPT-EDTA血清预处理以减少固相免疫测定（SPI）背景的功效。共纳入21份阴性对照(NC)值≥500的血清样品和20份NC值正常的血清样品，以评估潜在的不良反应。在SPI中25%的我们的样品中注意到高背景的问题。我们观察到SPT-EDTA血清预处理降低NC值<500的有效性为80％，与72％相比，ADS和SC处理的血清为67%，分别。有趣的是,我们发现SPT与ADS;ADS与SC处理血清在表型和单抗原珠测定中的抗体结合水平之间存在强相关性(p<0.001).对NC无不良影响，阳性对照（PC）值，与单独使用EDTA相比,预先使用SPT-EDTA的PC/NC比率。我们的数据显示，联合SPT-EDTA处理的血清比ADS更有效，和SC在降低SPI中的高背景。一起来看,在LuminexHLAAb测试之前的SPT-EDTA血清治疗具有成本效益，我们的实验室为Ab测试节省了近30%的年度总成本，并将测试周转时间提高了两个工作日。
