【作者】 刘忠；

【导师】 兰炯采； 高大勇；

【作者基本信息】 第一军医大学 ， 免疫学， 2004， 硕士

【摘要】 目的： 建立HPA基因型检测方法，探讨临床因HPA引起的血小板输注无效症的解决方案。 方法： 1、针对不同型别HPA基因间的差异，设计5对特异性引物，采用PCR-SSP方法，检测HPA(1-5)基因型： 2、HPA抗体筛选及鉴定采用微柱凝胶法及Capture-P方法： 3、对合肥地区的献血人群进行了HPA基因多态性调查，并建立已知HPA型的血小板供者登记库； 4、对多次输注血小板效果不良的患者进行了抗体筛选，并对因HPA抗体引起的血小板输注无效症等患者输注HPA基因型相合的血小板。 结果： 1、采用PCR-SSP方法检测HPA(1-5)基因型，敏感性强，特异性高，重复性好。 2、合肥地区献血人群HPA(1-5)基因中，HPA-4a 基因频率最高(0.993)，其次为HPA-1a(0.973)，HPA-4b基因频率最低(0.007)，其次是HPA-1b(0.027)；同时，建立了一支150名献血者组成的已知HPA型血小板供者资料库。 3、在32例血小板输注无效者中，6例检定为HPA抗体引起，其中4例为HPA-1a抗体，2例为HPA-5b抗体； 4、对6例HPA抗体引起的血小板输注无效者，在血小板供者库中选择相合的血小板输注，取得良好效果；结论: 采用PCR一SSP技术检测H以(1一5)基因型，在相同的扩增条件下，一次扩增可检测出H以(1一5)全部基因型，敏感性高，特异性强，重复性好;对献血员的H以基因多态性调查可以建立血小板供者库，能够有效地解决因H以抗体引起的血小板输注无效。更多还原

【Abstract】 Objective:To established HPA genotyping method and explored platelet transfusion refractoriness.Methods:1. According to difference of HPA gene, 5 pairs of oligonucleotide specific primers for HPA gene were designed and composed. The polymerase chain reaction sequence specific oligonucleotide primer (PCR-SSP) was used to amplify and detect the HPA gene.2. The methods of microcolumn gel test and Capture-p were used in screening and identification of HPA antibodies.3. HPA gene polymorphism in Hefei’s donor was investigated and platelet donor regirster was founded.4. Platelet-reactive antibodies in patients of platelet transfusion refractoriness were detected, and compatible donors were selected for platelet transfusion.Result:1. The method of HPA genotyping by PCR-SSP had high sensitivity .specificity and repetition.2. The highest frequencies of HPA genes was HPA -4a (0.993) in Hefei’s donor and the second was HPA-la (0.973). The lowest frequencies of HPA genes was HPA -4b (0.007) in Hefei’s donor, and the second was HPA-lb (0.027). At the same time, we established the platelets donor register.3. In 32 cases of platelet transfusion refractoriness, 6 cases resulted from HPA antibody. Of them, 4 cases were HPA-la, 2 case was HPA-5b.-4-4. Present study showed that platelets transfusion was successful by selecting HPA compatible donors.Conclusion:A method of genotyping was developed for HPA-1, -2, -3 and -5 by means of the PCR amplification with sequence-specific primers (PCR-SSP) technique. Primer sets were designed to allow PCR amplification for all systems using the same assay conditions. Specificity, sensitivity and repetition of the method are good.The HPA register can be established by investigation of HPA polymorphism in Hefei’s donor and it can overcome PTR that result from HPA antibodies.更多还原