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Abstract：
BACKGROUND: HLA antibodies might contribute to the pathogenesis of transfusion-related acute lung injury (TRALI). HLA antibody detection methods include ELISA, flow cytometry, and multiplex bead-based assays, as well as the older lymphocytotoxicity assay, and it is not obvious how to compare results across platforms.
METHODS: Five hundred twenty-five serum samples were selected from 7841 donors in the Leukocyte Antibody Prevalence Study (LAPS) repository based on risk for the development of HLA antibodies, using the number of pregnancies as the risk factor. Subjects included 81 males and females with 0 (n = 187), 1 (n = 67), or 2+ pregnancies (n = 190). Replicate frozen serum aliquots were sent blinded to four different HLA antibody assay manufacturers for detection using five different assays.
RESULTS: The flow cytometry and multiplex bead based-assays typically resulted in a larger proportion of HLA antibody positive samples compared with ELISA based assays. Latent variable analysis was used to derive a new set of consensus cutoffs, which yielded similar sensitivities across test platforms and increased concordance amongst assays. Assay agreement was higher in ever pregnant females than in males and never-pregnant females.
CONCLUSIONS: Different assays resulted in varied positivity rates when the manufacturer\'s suggested cutoffs were used, demonstrating that care needs to be taken when comparing clinical outcomes data generated using different HLA antibody assays and testing platforms. The method used here, involving latent variable analysis, presents one possible approach to calculating comparable cutoffs that result in broad agreement across assays with respect to positivity designation.
METHODS: Five hundred twenty-five serum samples were selected from 7841 donors in the Leukocyte Antibody Prevalence Study (LAPS) repository based on risk for the development of HLA antibodies, using the number of pregnancies as the risk factor. Subjects included 81 males and females with 0 (n = 187), 1 (n = 67), or 2+ pregnancies (n = 190). Replicate frozen serum aliquots were sent blinded to four different HLA antibody assay manufacturers for detection using five different assays.
RESULTS: The flow cytometry and multiplex bead based-assays typically resulted in a larger proportion of HLA antibody positive samples compared with ELISA based assays. Latent variable analysis was used to derive a new set of consensus cutoffs, which yielded similar sensitivities across test platforms and increased concordance amongst assays. Assay agreement was higher in ever pregnant females than in males and never-pregnant females.
CONCLUSIONS: Different assays resulted in varied positivity rates when the manufacturer\'s suggested cutoffs were used, demonstrating that care needs to be taken when comparing clinical outcomes data generated using different HLA antibody assays and testing platforms. The method used here, involving latent variable analysis, presents one possible approach to calculating comparable cutoffs that result in broad agreement across assays with respect to positivity designation.
摘要：
背景：HLA抗体可能与输血相关性急性肺损伤（TRALI）的发病机制有关。HLA抗体检测方法包括ELISA、流式细胞术,和多重基于珠子的检测，以及旧的淋巴细胞毒性试验，如何跨平台比较结果并不明显。
方法：根据HLA抗体发展的风险，从白细胞抗体患病率研究（LAPS）库中的7841名供体中选择了525份血清样品，使用怀孕次数作为危险因素。受试者包括81名男性和女性，其中0（n=187），1（n=67），或2+怀孕（n=190）。将复制的冷冻血清等分试样发送给四个不同的HLA抗体测定制造商,用于使用五个不同的测定进行检测。
结果:与基于ELISA的测定相比,流式细胞术和基于多重珠的测定通常导致更大比例的HLA抗体阳性样品。潜在变量分析用于得出一组新的共识截止值，这在测试平台上产生了相似的灵敏度，并增加了测定之间的一致性。曾经怀孕的女性的测定一致性高于男性和从未怀孕的女性。
结论：当使用制造商建议的截止值时，不同的测定导致不同的阳性率，证明在比较使用不同HLA抗体分析和测试平台产生的临床结局数据时,需要小心.这里使用的方法，涉及潜在变量分析，提出了一种可能的方法来计算可比较的截止值，该方法导致在阳性指定方面的测定之间达成广泛一致。
方法：根据HLA抗体发展的风险，从白细胞抗体患病率研究（LAPS）库中的7841名供体中选择了525份血清样品，使用怀孕次数作为危险因素。受试者包括81名男性和女性，其中0（n=187），1（n=67），或2+怀孕（n=190）。将复制的冷冻血清等分试样发送给四个不同的HLA抗体测定制造商,用于使用五个不同的测定进行检测。
结果:与基于ELISA的测定相比,流式细胞术和基于多重珠的测定通常导致更大比例的HLA抗体阳性样品。潜在变量分析用于得出一组新的共识截止值，这在测试平台上产生了相似的灵敏度，并增加了测定之间的一致性。曾经怀孕的女性的测定一致性高于男性和从未怀孕的女性。
结论：当使用制造商建议的截止值时，不同的测定导致不同的阳性率，证明在比较使用不同HLA抗体分析和测试平台产生的临床结局数据时,需要小心.这里使用的方法，涉及潜在变量分析，提出了一种可能的方法来计算可比较的截止值，该方法导致在阳性指定方面的测定之间达成广泛一致。
