a Shanghai Institute of Blood Transfusion , Shanghai Blood Center , Shanghai , China.
Platelets. 2018 Jan;29(1):71-75. doi: 10.1080/09537104.2017.1294681. Epub 2017 Apr 13.
Human platelet antigens (HPAs) are platelet-specific alloantigens that cause alloimmunization. Alloantibodies induced via pregnancy and transfusion may result in various clinical syndromes. The monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay is the gold standard for the detection and identification of HPA antibodies. The present study aimed to develop standardized quantitative MAIPA assays for antibodies against HPA-1a, 3a, and 5b. MAIPA protocol was applied on serial dilutions of standard or reference reagents containing antibodies against HPA-1a, 3a, and 5b. Optimal experimental parameters were determined, and standard curves were constructed with a linear regression. The sensitivity, specificity, and reproducibility of the assays were also evaluated. The optimum reagents and conditions for the MAIPA assays were decided through repeated testing and comparison. The intra-assay coefficients of variation (CVs) are 1.33.6%, 2.75.2%, and 4.75.3%; the inter-assay CVs are 4.05.6%, 4.45.9%, and 3.26.3%, respectively, for HPA-1a, HPA-3a, and HPA-5b antibodies. In specificity tests, some monoclonal antibodies and sera with other antibodies were tested and no obvious positive result was observed. The standardized quantitative MAIPA assays for antibodies against HPA-1a, 3a, and 5b have good sensitivity, reproducibility, and specificity. They have important application value in clinical diagnosis of alloimmunization disease caused by HPA antibodies and safe transfusion of platelets.
人类血小板抗原（HPAs）是引起同种免疫的血小板特异性同种抗原。通过妊娠和输血产生的同种抗体可能导致各种临床综合征。单克隆抗体特异性血小板抗原固定（MAIPA）检测法是检测和鉴定 HPA 抗体的金标准。本研究旨在开发针对 HPA-1a、3a 和 5b 抗体的标准化定量 MAIPA 检测法。MAIPA 方案应用于含有针对 HPA-1a、3a 和 5b 抗体的标准或参考试剂的连续稀释液。确定了最佳实验参数，并通过线性回归构建了标准曲线。还评估了检测法的灵敏度、特异性和重现性。通过反复测试和比较确定了 MAIPA 检测法的最佳试剂和条件。MAIPA 检测法的批内变异系数（CV）分别为 1.3%3.6%、2.7%5.2%和 4.7%5.3%；批间 CV 分别为 4.0%5.6%、4.4%5.9%和 3.2%6.3%，分别用于 HPA-1a、HPA-3a 和 HPA-5b 抗体。在特异性测试中，测试了一些单克隆抗体和含有其他抗体的血清，未观察到明显的阳性结果。针对 HPA-1a、3a 和 5b 抗体的标准化定量 MAIPA 检测法具有良好的灵敏度、重现性和特异性。它们在 HPA 抗体引起的同种免疫性疾病的临床诊断和血小板安全输血方面具有重要的应用价值。
