Costa Thiago Henrique, Bonet-Bub Carolina, Kutner José Mauro
Hospital Israelita Albert Einstein, São Paulo, SP, Brazil.
Hospital Israelita Albert Einstein, São Paulo, SP, Brazil.
Hematol Transfus Cell Ther. 2024 Nov;46 Suppl 5(Suppl 5):S97-S102. doi: 10.1016/j.htct.2024.06.004. Epub 2024 Sep 7.
The identification of platelet antibodies is essential for diagnosing and managing conditions such as fetal and neonatal alloimmune thrombocytopenic purpura, post-transfusion purpura, and immune platelet refractoriness. Monoclonal antibody immobilization of platelet antigens (MAIPA) is the standard method for detecting anti-human platelet antigen (HPA) antibodies, while the detection of anti-HLA antibodies once relied on the complement-dependent cytotoxicity method, however advanced technologies such as enzyme-linked immunosorbent assay and Luminex have significantly improved sensitivity and accuracy in identifying these antibodies. Flow cytometry-based techniques (platelet immunofluorescence test - PIFT) and Luminex platform-driven microsphere-based multiplex assays (Pak-Lx) are widely employed in platelet immunology laboratories owing to their remarkable flexibility and versatility. The present study compared the sensitivity, specificity, and concordance of these different serological techniques used in platelet antibody identification.
One hundred serum samples from patients suspected of immune-mediated platelet disorders were examined. Initially, the samples underwent testing using the MAIPA method. Subsequently, the results were compared with three alternative methods: PIFT and microsphere-based multiplex assays for both HLA and HPA antibodies.
Pak-Lx demonstrated a 94 % agreement with MAIPA, while PIFT had 88 % agreement for HPA antibodies. For HLA antibody detection, Pak-Lx versus DLX had 75 % concordance, MAIPA versus DLX showed 77 %, and PIFT versus DLX displayed an 81 % concordance rate. Remarkably, there were no significant differences in concordance levels between Pak-Lx and PIFT compared to MAIPA and DLX for anti-HPA and HLA antibodies, respectively.
This study found no significant differences in concordance among the tested assays for detecting anti-HPA and anti-HLA antibodies. These data suggest that no single method can detect all clinically important antibodies. Therefore, it is advisable that each laboratory develops customized protocols based on their expertise and employs complementary methods for comprehensive patient assessments.
血小板抗体的鉴定对于诊断和管理诸如胎儿及新生儿同种免疫性血小板减少性紫癜、输血后紫癜和免疫性血小板输注无效等病症至关重要。血小板抗原单克隆抗体固定法（MAIPA）是检测抗人血小板抗原（HPA）抗体的标准方法，而抗HLA抗体的检测曾依赖补体依赖细胞毒性法，不过酶联免疫吸附测定和Luminex等先进技术在鉴定这些抗体方面显著提高了灵敏度和准确性。基于流式细胞术的技术（血小板免疫荧光试验 - PIFT）和Luminex平台驱动的基于微球的多重分析（Pak-Lx）因其卓越的灵活性和通用性而在血小板免疫学实验室中广泛应用。本研究比较了用于血小板抗体鉴定的这些不同血清学技术的灵敏度、特异性和一致性。
检测了100份疑似免疫介导血小板疾病患者的血清样本。首先，使用MAIPA方法对样本进行检测。随后，将结果与三种替代方法进行比较：针对HLA和HPA抗体的PIFT和基于微球的多重分析。
Pak-Lx与MAIPA的一致性为94%，而PIFT对HPA抗体的一致性为88%。对于HLA抗体检测，Pak-Lx与DLX的一致性为75%，MAIPA与DLX为77%，PIFT与DLX的一致性率为81%。值得注意的是，与MAIPA和DLX相比，Pak-Lx和PIFT在抗HPA和抗HLA抗体的一致性水平上分别没有显著差异。
本研究发现，在检测抗HPA和抗HLA抗体的测试方法之间，一致性没有显著差异。这些数据表明，没有单一方法能够检测出所有临床上重要的抗体。因此，建议每个实验室根据自身专业知识制定定制方案，并采用互补方法进行全面的患者评估。
