作者：NguyenD ,GobelM ,SchoberM ,KluterH ,PanzerS

德国曼海姆，黑森Baden - Wurttemberg红十字血液服务中心

奥地利维也纳大学医学院， 血型血清学实验室及输血科

来源: Vox Sanguinis, 2010 july, Supplement1,Vol 99

背景：各同种抗体的检测和特异性检测，对同种免疫血小板减少症有重要的诊断意义，如新生儿同种免疫血小板减少症以及血小板输注无效，另一方面，自身抗体的检测也有助于自身免疫血小板减少症的诊断。同种抗体和自身抗体的靶组织可能是不同的血小板糖蛋白。同步分析血小板特殊抗体（SASPA）检测的基于用流式细胞仪技术对不同的血小板特殊抗体的同步检测，并已在我们曼海姆研究所成为常规应用。在本研究中，我们采用SASPA与“金标准”MAIPA试验相比较，进行了对血小板特殊抗体检测的盲比研究。

方法：194个有或没有潜在性疾病病人的血清，这些血清来源于自送检血小板抗体。血清由曼海姆和维也纳间交换（99份来自曼海姆，95份来自维也纳）。全部194个血清标本在维也纳用MAIPA法检测，在曼海姆则用SASPA法检测。怀疑有同种抗体或自身抗体的血清标本为随机抽取，调查抗GPⅡb/Ⅲa, GPⅠb/Ⅸ, GPⅠa/Ⅱa, GPⅣ的血小板特殊抗体以及血小板反应性HLA Ⅰ类抗体。每个实验室按它们各自常规应用所建立的程序，使用试验用血小板和单克隆鼠抗体（mAb）。所有报告的数据，对各自其它试验方法所得的数据都是双盲的。灵敏度研究包括针对HPA1a,-1b,-3b,-5b,-15b 和 HLA Ⅰ类已知抗体的稀释试验。

结果：总体而言，153份（78.9%）血清标本的结果是一致的。用MAIPA法或SASPA法检测出的、或两种方法都检测出的所有的同种抗体，与病人的基因型和临床表现上是符合的。如果以MAIPA方法的结果为参照，SASPA方法检测同种抗体的特异性和灵敏度分别为97.3%和86.3%，检测自身抗体的特异性和灵敏度分别为95.3%和44.9%。倍比稀释实验的比较：较MAIPA法而言，SASPA法可在更低浓度中检测到同种抗体。

结论：在此首次实验室之间盲比试验中显示，SASPA法可适用于血小板抗体的检测，特异性和灵敏度与金标准MAIPA法相似。SASPA法的优点在于可同步检测抗体，试验中所需要的血小板量更少，以及较高的灵敏度，特别是在检测同种抗体的情况下。

DETECTION AND DIFFERENTIATION OF PLATELET ALLO- AND AUTO-ANTIBODIES USING SASPA AND MAIPA: A BLIND INTERLABORATORY COMPARISON

NguyenD ,GobelM ,SchoberM ,KluterH ,PanzerS

Vox Sanguinis, 2010 july, Supplement1,Vol 99

Background: The detection and specification>Mannheim institute. In this study, we performed a blind interlaboratory comparative investigation of platelet-specific antibodies using SASPA versus the "gold standard", the MAIPA assay.

Methods: Sera came from 194 patients with and without various underlying disorders. The sera were submitted for the detection of platelet antibodies. An exchange of sera took place between Mannheim and Vienna (99 sera from Mannheim, 95 sera from Vienna). All samples were tested by the MAIPA assay in Vienna, and by the SASPA assay in Mannheim. Sera with suspected platelet allo- or autoantibodies were randomly selected for the investigation on platelet-specific antibodies against GPⅡb/Ⅲa, GPⅠb/Ⅸ, GPⅠa/Ⅱa, GPⅣ and platelet reactive HLA classⅠ. Each laboratory used test platelets and monoclonal mouse antibodies (mAb) according to the procedures established for their routine use. All data were reported blinded to those from the respective other method. Sensitivity studies included dilution studies with known antibodies against HPA1a, -1b, -3b, -5b, and -15b and HLA class Ⅰ.

Results: Overall, results were concordant for 153 sera (78.9%). All detected alloantibodies, either with MAIPA or SASPA method or both methods were compatible with the patients’ genotypes and the clinical observations. The specificity and sensitivity of SASPA, based on the MAIPA results, were 97.3% and 86.3%, respectively, for the detection of alloantibodies. The respective results for the detection of  autoantibodies were 95.3% and 44.9%. In serial dilution experiments, all alloantibodies were detectable at lower concentrations by SASPA than by MAIPA.

Conclusions: In this first blind interlaboratory-comparison, the SASPA assay was shown to be a suitable method for detecting platelet antibodies and similar to the gold standard MAIPA with regard to sensitivity and specificity. The advantages of SASPA are the simultaneous detection of antibodies, the need of less platelets to perform this assay and the higher sensitivity, particularly with alloantibodies.

本栏目负责人：黎海燕