作者：Chong W, Metcalfe P, Mushens R, Lucas G, Ouwehand WH, Navarrete CV.

来源：Transfusion. 2010 Dec 16.

摘要 背景：人类血小板抗原(HPAs)同种抗体的检测，对临床诊断胎母同种免疫血小板减少症(FMAIT)、输血后紫癜及血小板输注无效是很重要的。目前，大部分血小板同种抗体的检测方法依赖于已知HPA分型的血小板谱细胞，一些还依赖血小板糖蛋白单克隆抗体(MoAbs)。具有HPA-1a(rHPA-1a)或HPA-1b(rHPA-1b)表位的重组β3整合素片段，已经被生产来作为可供选择的抗原来源。本文应用Luminex xMAP技术，来评估这些整合素片段是否适合HPA-1a同种抗体检测。

实验设计与方法：3-plex微球是由生物素标记的rHPA-1a，rHPA-1b和重组糖蛋白VI连结到LumAvidin微球上。40个已经用MAIPA法诊断的FMAIT病人标本，用来评估本试验。

结果：用rHPA-1a-及rHPA-1b-连结的微球，能检测到所有含有HPA-1特异抗体病人标本中的HPA-1a和HPA-1b同种抗体。而且，此连结的微球与HLA-I类抗体无交叉反应。

结论：该用来检测临床FMAIT病人HPA-1a抗体的3-plex微球具有较高的灵敏度和特异性。其它重组整合素片段的发展和Luminex xMAP技术的使用，将有助于HPA抗体的快速检测及血小板同种免疫紊乱的快速诊断。

Detection of human platelet antigen-1a alloantibodies in cases of fetomaternal alloimmune thrombocytopenia using recombinant β3 integrin fragments coupled to fluorescently labeled beads

Chong W, Metcalfe P, Mushens R, Lucas G, Ouwehand WH, Navarrete CV.

Transfusion. 2010 Dec 16.

Abstract BACKGROUND: Testing for alloantibodies against human platelet antigens (HPAs) is essential for the clinical diagnosis of fetomaternal alloimmune thrombocytopenia (FMAIT), posttransfusion purpura, and platelet (PLT) refractoriness. Most of the methods currently used for HPA alloantibody detection rely on the availability of panels of HPA-typed PLTs and some rely on validated monoclonal antibodies (MoAbs) against the PLT glycoproteins. Recombinant β3 integrins displaying the HPA-1a (rHPA-1a) or HPA-1b (rHPA-1b) epitopes have been produced as an alternative source of antigen. The suitability of these integrin fragments was evaluated for the development of an HPA-1a alloantibody screening assay, using Luminex xMAP technology.

STUDY DESIGN AND METHODS: A 3-plex bead assay was developed by coupling biotinylated rHPA-1a, rHPA-1b, and recombinant glycoprotein VI to LumAvidin microspheres. Forty patient samples referred for FMAIT diagnostic testing, which were previously screened by the MoAb-specific immobilization of PLT antigens (MAIPA) assay, were used to assess the assay.

RESULTS: The rHPA-1a- and rHPA-1b-coupled beads were able to detect HPA-1a and HPA-1b alloantibodies in all patient samples tested that were previously confirmed to contain HPA-1-specific antibodies. Furthermore, HLA Class I antibodies did not cross-react with the coupled beads.

CONCLUSION: The 3-plex bead assay can be used to detect HPA-1a antibodies with sufficient specificity and sensitivity for use in the clinical setting of FMAIT. The development of other recombinant integrin fragments with the use of Luminex xMAP technology may assist in providing more rapid HPA antibody detection, enabling prompt diagnosis of alloimmune PLT disorders.

翻译：钟周琳 校对：裴永峰