限制性片段长度多态性分析人血小板抗原5系统基因型

Genotyping of human platelet antigen system 5 by PCR restriction fragment length polymorphism

目的 建立人类血小板抗原 (HPA) 5系统限制性片段长度多态性 (PCR RFLP)分析方法。方法 采用PCR RFLP方法对 2 5名健康献血员的HPA 5系统进行基因分型。分型结果与采用等位基因特异性寡核苷酸点杂交 (PCR ASO)获得的结果进行比较。结果 以PCR RFLP方法对 2 5名献血员的HPA 5系统分型 :2 4名为a/a型 ,1名为a/b型 ,未发现b/b纯合子个体 ,结果与PCR ASO方法获得的结果完全一致。结论 使用PCR RFLP方法对HPA 5系统进行基因分型 ,具有简便、快速、准确等优点 ,有利于新生儿同种免疫血小板减少性紫癜。

Objective To develop a polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) method using designed primers for determining the genotype of humen platelet antigens (HPA)5 system. Methods HPA 5 system of 25 healthy blood donors were genotyped using PCR RFLP method. The results obtained by PCR RFLP were compared with those determined by allele specific oligonucleotid hybridization (PCR ASO). Results The results of HPA 5 system obtained by PCR RFLP in 25 health donors were as follows: 24 of aa, 1 of ab and 0 of bb. All were in good agreement with those determined by PCR ASO. Conclusions Because PCR RFLP method is plain, fast and reliable for HPA 5 system genotyping, it is suitable for the diagnosis and therapy of neonatal alloimmune thrombocytopenia, posttransfusion purpura, platelet transfusion refractoriness and so on..