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目的分析中国南方人群HPA-15基因型分布频率,研究血小板上HPA-15抗原表达量。寻找较好的血小板保存方法以维持HPA-15的高表达量,为单克隆抗体俘获血小板抗原技术(MAIPA)检测HPA-15同种抗体提供合适的供者血小板。方法 1)随机选取广州血液中心的健康血小板单采献血者109名,留取EDTA抗凝管全血。SSPPCR方法检测HPA-15基因型。流式细胞术检测血小板上HPA-15抗原表达量。2)另随机选取健康献血者的单采血小板67例,将每份血小板分为2份,1份重悬于EDTA-PBS中放于4℃保存作为检测组A,另1份留于血小板辫子中放于血小板振荡仪中作为检测组B,在d 0、1、2、3、7、14检测2组血小板上HPA-15的检测含量,比较2组HPA-15含量的变化。结果 1)109名随机健康献血者HPA-15a和HPA-15b的基因频率是0.495和0.505。流式细胞术检测HPA-15抗原的MFI值的范围2.44-20.4,阳性率3.2%-86.7%。不同HPA-15基因型之间,HPA-15抗原表达量有统计学差异(P<0.05)。2)分析比较AB这2个组,随着测量时间点的推移,2组HPA-15的表达量均有降低,但检测组B中HPA-15的表达含量高于检测组A,且2组的MFI值有统计学差异(P<0.05)。结论 HPA-15a和HPA-15b的基因分布频率相近,HPA-15抗原表达含量整体水平偏低且个体差异性大。体外保存方式下,HPA-15的表达含量随着时间推移逐渐降低,4℃保存方式下HPA-15含量降解尤为明显。血小板振荡仪中保存血小板的方法为MAIPA检测HPA-15抗体提供更合适的血小板抗原。
Objective To analyze the distribution frequency of HPA-15 genotypes in southern Chinese population and to study the expression of HPA-15 antigen on platelets. To find a better method of platelet preservation in order to maintain the high expression of HPA-15 and to provide appropriate donor platelets for monoclonal antibody capture platelet antigen technology (MAIPA) detection of HPA-15 allo-antibodies. Methods 1) Randomly selected 109 healthy blood platelet-collecting blood donors from Guangzhou Blood Center and collected EDTA anticoagulant whole blood. SSPPCR method to detect HPA-15 genotype. Flow cytometry was used to detect the expression of HPA-15 antigen on platelets. 2) Randomly select 67 healthy apheresis platelets from healthy blood donors, divide each platelet into 2 parts, 1 part resuspend in EDTA-PBS and store at 4 ° C. as test group A, and the other 1 part remain in platelet braid In the platelet shaker as a test group B, on the d 0,1,2,3,7,14 detection of two groups of platelet HPA-15 detection levels were compared two groups of HPA-15 content changes. Results 1) The gene frequencies of HPA-15a and HPA-15b in 109 randomly selected healthy donors were 0.495 and 0.505, respectively. The range of MFI value of HPA-15 antigen detected by flow cytometry was 2.44-20.4, the positive rate was 3.2% -86.7%. There was a significant difference in HPA-15 antigen expression between different HPA-15 genotypes (P <0.05). 2) Compared with the two AB groups, the expression of HPA-15 in both groups decreased as time went by, but the content of HPA-15 in group B was higher than that in group A The MFI values were statistically different (P <0.05). Conclusion The frequency of HPA-15a and HPA-15b gene distribution is similar, the overall level of HPA-15 antigen expression is low and the individual differences are large. In vitro preservation, HPA-15 expression levels gradually decreased over time, 4 ℃ preservation mode HPA-15 content degradation is particularly evident. The method of platelet preservation in platelet oscillators provides a more suitable platelet antigen for MAIPA detection of HPA-15 antibodies.