摘要
目的:研究血小板膜糖蛋白Ⅱb(GPIIb)三种基因多态性之间的关系及GPIIbT13959G在血小板输注耐受中的作用。方法:聚合酶链反应-单链构型多态性分析检测110例正常人GPIIb基因第26和30外显子(Exon26,Exon30)及21内含子(Intron21)基因多态性,进行基因序列分析,研究这些基因多态性是否存在连锁关系;应用Fok1酶切法对147例血液病人人类血小板抗原-3(humanplateletantigen-3,HPA-3)基因进行分型,并与110例正常人进行比较;将接受单采血小板输注的44例血液病人随机分为HPA-3同型输注组和对照组,血小板输注3次以后检测血小板抗体。结果:正常人GPIIb存在基因多态性,表现分别为gDNA13959位点T→G,16977位点C→T,11996～12004位点9个碱基缺失,且三者具有同步性。Fox1酶切分析结果表明,正常人和血液病人的HPA-3a基因频率分别为83.6%(92/110)和81.9%(119/147),HPA-3b基因频率分别为16.4%(18/110)和19.1%(28/147),两者之间差异无统计学意义(P>0.05)。血液病人接受单采血小板输注后,HPA-3同型输注组血小板抗体阴性,而对照组有2例存在血小板抗体,其中1例为HPA-3a抗体。结论:(1)GPIIb基因存在gDNA13959位点T→G,16977位点C→T,11996～12004位点9个碱基缺失,这3个位点的多态性可能存在完全连锁关系;(2)血液病人与正常人的HPA-3基因频率相同;(3)HPA-3系统可能是我国人群产生血小板输注耐受的原因之一。
Objective To investigate the relationship among 3 polymorphisms of GP Ⅱb and the function of GP Ⅱb T13959 G in the platelet transfusion refractoriness （PTR）. Methods The 26th exon, the 30th exon and the 21st intron of gene GP Ⅱb in 110 individuals were amplified by polymerase chain reaction （PCR） , and the PCR products were analyzed with single-strand conformation polymorphism （SSCP） and sequenced to investigate whether there was linkage among the polymorphisms of the gene. Human platelet antigen-3 （ HPA-3 ） gene frequency was detected by Fok Ⅰ enzyme in 147 patients with hematologic diseases, and was compared with that in 110 normal individuals. Forty-four patients who received apheresis platelet transfusion repeatedly were randomly divided into the HPA-3 homotype group and the control group. The antibodies of the platelet were detected after 3 times of platelet transfusion. Results There were polymorphisms of gene GP Ⅱb in the 26th, 30th exon and the 21 st intron, and the mutations were： T changed into G in 13 959th of the 26th exon; C changed into T in 16 997th of the 30th exon; the 9 bps deletion occurred in 11 996 - 12 004th of the 21 st intron. The 3 polymorphisms had synchronization in the individuals. The results of Fok Ⅰ enzyme indicated that the frequency of HPA-3a was 83. 6% （92/110） and 81. 9% （ 119/147）, and that of HPA-3b was 16.4% （18/110） and 19. 1% （28/147） in the normal individuals and the patients respectively. There was no significant difference between the patients and normal individuals （ P 〉 0.05 ）. After the platelet transfusion, the antibodies of all the cases of the homotype platelet transfusion were negative, but the antibodies in 2 cases of the control group were positive, and there was antibody to HPA-3 a in one of the antibodies positive cases. Conclusion （ 1 ）There is close linkage among the polymorphisms of gene GP Ⅱb, which is T→G in 13 959th of the 26th exon, C→T in 16 997th of the 30th exon, and the 9 bps deletion in 11 996 N 12 004th in the 21 st intron. （2）The gene frequency of HPA-3a/3b is similar in the normal individuals and patients with hematologic diseases. （3） HPA-3 system may be one of the reasons for PTR in Chinese.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2007年第4期584-589,共6页
Journal of Central South University ：Medical Science
基金
卫生部临床学科重点项目(97080242)
湖南省卫生厅科研基金(B2005-071)~~
关键词
血小板
多态性
基因频率
同种抗原
输血
platelet
polymorphism
gene frequency
alloantigen
transfusion
