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A case report-Two manufacturers SAB testing kits can reveal different HLA antibody profiles-Identifying prozone and denatured antigen.
HLA ( IF 5.9 ) Pub Date : 2020-05-12 , DOI: 10.1111/tan.13913
Richard K Battle 1 , Lorna Henderson 2 , Paul J Phelan 2 , Katy Latham 3 , David M Turner 1
Affiliation  

Recent reports have identified that the presence of non‐native conformation HLA to which antibody can bind upon Luminex HLA Class I single antigen beads, can vary in levels between different manufacturers kits and that the prozone effect may also be specific to particular products. We present a case in which both prozone and non‐native HLA reactive antibodies were observed, which raises important questions on how SAB assays are utilised, especially in the post‐transplant monitoring setting. A 56‐year old, highly sensitised female patient awaiting a regraft received a HLAi renal transplant. Post‐transplant monitoring showed discordant results between two SAB manufacturers assays, with one assay identifying a potential de novo HLA DSA. HLA Class I antibody reactivity was observed which was directed towards the Bw6 public epitope, which is present upon HLA molecules encoded for by numerous HLA‐B alleles. However in the day 19 post‐transplant sample reactivity spread beyond the Bw6 epitope. To investigate the possibility of a prozone type effect influencing the testing kit the day 19 post‐transplant sample was diluted 1:10 with PBS and reanalysed. After dilution the Bw6 reactivity was observed again, however the suspect de novo DSA still persisted. An analysis of the mismatched epitopes identified one manufacturer's assay as being confounded by the presence of denatured reactivity as well as prozone.

中文翻译:

病例报告-两家制造商的SAB检测试剂盒可以揭示不同的HLA抗体谱-鉴定前带和变性抗原。

最近的报告已经确定,在Luminex HLA I类单抗原珠上可以结合抗体的非天然构象HLA的存在,在不同制造商试剂盒之间的水平可能有所不同,并且脯氨酸的作用也可能对特定产品具有特异性。我们提出了同时观察到prozone和非天然HLA反应性抗体的情况,这引发了关于如何利用SAB测定法的重要问题,尤其是在移植后监测环境中。一名等待移植的56岁,高度敏感的女性患者接受了HLAi肾移植。移植后监测显示,两种SAB制造商测定法之间的结果不一致,其中一种测定法确定了潜在的从头HLA DSA。观察到针对Bw6公共表位的HLA I类抗体反应性,存在于众多HLA-B等位基因编码的HLA分子上。但是,在移植后第19天,样品的反应性扩散到了Bw6表位之外。为了研究前区类型效应影响检测试剂盒的可能性,将移植后第19天的样品用PBS以1:10稀释并重新分析。稀释后,再次观察到Bw6反应性,但是可疑的从头DSA仍然存在。对错配表位的分析确定了一个制造商的检测方法被变性的反应性以及前带的存在所混淆。用PBS进行10次分析。稀释后,再次观察到Bw6反应性,但是可疑的从头DSA仍然存在。对不匹配的表位的分析确定了一个制造商的检测方法被变性的反应性和脯氨酸的存在所混淆。用PBS进行10次分析。稀释后,再次观察到Bw6反应性,但是可疑的从头DSA仍然存在。对不匹配的表位的分析确定了一个制造商的检测方法被变性的反应性和脯氨酸的存在所混淆。
更新日期:2020-05-12
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