槽式印迹法和流式细胞术：血小板抗体筛选的两种有效方法，用于血小板难治性患者。

Slot blotting and flow cytometry: two efficient assays for platelet antibody screening among patients with platelet refractoriness.

机构信息

Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.

Cell Therapy and Regenerative Medicine Comprehensive Center, Kerman University of Medical Sciences, Kerman, Iran.

出版信息

Vox Sang. 2021 Jan;116(1):106-115. doi: 10.1111/vox.12988. Epub 2020 Aug 8.

DOI:10.1111/vox.12988

PMID:32770852

Abstract

BACKGROUND AND OBJECTIVES

Frequent platelet transfusion may lead to the formation of alloantibodies and immune-mediated platelet destruction. Currently, identifying economic and effective screening methods is necessary for the management of platelet transfusion while different tests were recommended. The present study aims to challenge the performance of slot blotting (SB) and flow cytometry (FC) assays in detecting immune platelet refractoriness.

MATERIALS AND METHODS

Sera from 118 patients who received blood components and were clinically suspected of platelet refractoriness were enrolled. Platelet-reactive antibodies were explored in parallel by SB, FC and monoclonal antibody-specific immobilization of platelet antigens (MAIPA) techniques. In a further study, chloroquine-treated platelets were incubated with MAIPA-positive serum, and then, the results of the SB and FC techniques were compared.

RESULTS

Using MAIPA as a reference, antibodies were detected in 51 sera, with specificity for human leucocyte antigens (HLA), human platelet antigens (HPA) or both HLA/HPA, in 27, 18 and 6 patients, respectively. The sensitivity and specificity of SB and FC were 86·3%, 88·1%, 82·4% and 95·5%, respectively. The Spearman correlation revealed significant (P < 0·001) correlations between FC (r = 0·763) and SB (r = 0·738) with MAIPA. In respect to HPA antibody detection, SB had 83·3% sensitivity and 92·6% specificity compared to 91·7% and 96·3% for FC while both approaches are acceptable (P < 0·001, r = 0·69; P < 0·001, r = 0·773) and can be recommended.

CONCLUSIONS

The present study acknowledges that among the used methods, the flow cytometry's performance is the most appropriate, but slot blotting, with acceptable sensitivity, can be used as an acceptable and convenient procedure for platelet antibody screening.

摘要

背景与目的

频繁的血小板输注可能导致同种异体抗体的形成和免疫介导的血小板破坏。目前，在管理血小板输注时，需要确定经济有效的筛选方法，同时推荐不同的检测方法。本研究旨在比较斑点印迹（SB）和流式细胞术（FC）检测免疫性血小板反应性的效果。

材料与方法

收集了 118 例接受血液成分并临床怀疑血小板反应性的患者的血清。同时平行采用 SB、FC 和单克隆抗体特异性血小板抗原固定（MAIPA）技术检测血小板反应性抗体。在进一步的研究中，将氯喹处理的血小板与 MAIPA 阳性血清孵育，然后比较 SB 和 FC 技术的结果。

结果

以 MAIPA 为参考，在 51 份血清中检测到抗体，其中 27、18 和 6 例患者的抗体特异性分别为人白细胞抗原（HLA）、人类血小板抗原（HPA）或 HLA/HPA 均阳性。SB 和 FC 的敏感性和特异性分别为 86.3%、88.1%、82.4%和 95.5%。Spearman 相关分析显示，FC（r=0.763）和 SB（r=0.738）与 MAIPA 之间存在显著相关性（P<0.001）。在 HPA 抗体检测方面，SB 的敏感性为 83.3%，特异性为 92.6%，FC 的敏感性为 91.7%，特异性为 96.3%，两者均具有可接受性（P<0.001，r=0.69；P<0.001，r=0.773），均可推荐使用。