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1.
吉林地区汉族人群血小板抗原系统基因分型研究   总被引:1,自引:0,他引:1  
张冬霞 《中国输血杂志》2011,24(10):869-870
目的研究吉林省汉族人群人血小板同种抗原系统(HPA)基因及其多态性分布特征,建立已知HPA基因型的血小板捐献者资料库。方法用DNA试剂盒提取外周血标本中DNA,通过特异性引物(PCR-SSP)扩增HPA等位基因。检测200名HPA1-6,15抗原系统共14个抗原的基因分型。结果 200名健康、无血缘关系的吉林省汉族人群HPA基因频率为:HPA-1a0.9900、-1b0.0100;-2a0.9300、-2b0.0700;-3a0.5575、-3b0.4425;-4a1.0000、-4b0;-5a0.9900、-5b0.0100;-6a0.9875、-6b0.0125;-15a0.5125、-15b0.4875。HPA-3与我国其它地区同类资料比较χ2=3.18,P>0.05;与亚洲印尼、泰国、英国及刚果相比χ2=5.98,P>0.05;HPA-5与HPA-15和刚果、喀麦隆比较χ2=55.06、χ2=15.63,P<0.01结论吉林地区血小板志愿捐献者HPA-1~6和HPA-15抗原系统的基因频率符合Hardy-Wein-berg遗传定律,与其它地区和国家的同类资料相比显示出种族和地域性差异,由此建立起了吉林地区已知HPA基因型的血小板捐献者资料库。  相似文献   

2.
新疆汉族人群血小板抗原1—5、15系统基因多态性分析   总被引:2,自引:0,他引:2  
目的调查研究与血小板输注无效关系密切的人类血小板抗原(human platelet antigen,HPA)1—5及15系统基因在新疆汉族人群中的遗传多态性。方法采用聚合酶链-序列特异性引物(PCR-SSP)法对101例无血缘关系的新疆汉族血样进行HPA基因分型。结果新疆汉族HPA-1a、2a、3a、4a、5a、15a基因频率分别是0.9851、0.9208、0.5446、1、0.9505、0.4653,HPA-1b、2b、3b、4b、5b、15b基因频率分别是0.0149、0.0792、0.4554、0、0.0495、0.5347,新疆汉族HPA基因频率与维吾尔族人群相比,HPA-1a、1b基因频率差异具有统计学意义(P<0.05)。结论HPA-1、2、3、5、15系统均具有多态性,HPA-3、15系统具有高度多态性。  相似文献   

3.
摘要:目的:研究河南地区汉族人群血小板抗原基因分布频率,建立地区性血小板抗原基因库,为血小板配合输注提供实验依据。 方法:用序列特异性引物聚合酶链反应(PCR-SSP)对160例无血缘关系的河南汉族体检健康者进行HPA1-16bw系统基因分型。 结果:HPA1-6和HPA-15基因频率分别为1a 0.987 5,1b 0.012 5; 2a 0.946 9,2b 0.053 1; 3a 0.590 6,3b 0.409 4; 4a 0.996 9,4b 0.003 1; 5a 0.990 6,5b 0.009 4; 6a 0.981 2,6b 0.018 8;15a 0.540 6,15b 0.459 4。 HPA7-14bw,16bw仅检测到a/a等位基因,基因频率均为1.000 0。经χ2检验各系统HPA符合Hardy-Weinberg平衡法则。HPA基因分布存在种族和地区差异。 结论:河南汉族人HPA-15 和HPA-3基因型杂合率最高。HPA基因分型有助于输注血小板时降低免疫性血小板减少症的发生概率。  相似文献   

4.
目的检测安徽合肥地区单采血小板捐献者血样的HPA基因分型,统计本地区血小板捐献者HPA基因型分布频率。方法①采用多重荧光PCR的方法对1181名18-55周岁的单采血小板捐献者血样进行HPA1-6,10,15,21的基因分型检测。②采用SBT直接测序的方法对22名18-55周岁的单采血小板捐献者血样进行HPA基因型分型检测。结果发现安徽地区人群中HPA-2,HPA-3,HPA-15最具有多态性,HPA各等位基因型频率分别为:1a∶0.9941,1b∶0.0059,2a∶0.9462,2b∶0.0538,3a∶0.5741,3b∶0.4259,4a∶0.9987,4b∶0.0013,5a∶0.9818,5b∶0.0182,6a∶0.9852,6b∶0.0148,10a∶1.0,10b∶0.0,15a∶0.5394、15b∶0.4606,21a∶0.9928、21b∶0.0072。结论使用多重荧光方法与SBT直接测序方法HPA基因分型检测结果一致性较好;安徽合肥地区人群的HPA基因分型的多态性与文献中报道的其他地区基本一致,但基因型分布频率有本地区的特点。  相似文献   

5.
目的对辽宁地区血小板志愿捐献者进行HPA基因分型,建立已知HPA基因型的血小板捐献者资料库。方法采用PCR-SSP方法对随机抽取的130名血小板捐献者进行HPA-1-6和HPA-15抗原系统共14个抗原的基因分型。结果辽宁地区血小板志愿捐献者HPA的基因频率分别是:HPA-1a0.9925、1b0.0075、2a0.9305、2b0.0695、3a0.5810、3b0.4190、4a1.0000、4b0.000、5a0.9810、5b0.0190、6a0.9885、6b0.0115、15a0.4615、15b0.5385。结论辽宁地区血小板志愿捐献者HPA-1—6和HPA-15抗原系统的基因频率符合Hardy-Wein-berg遗传定律,与其它地区和国家的同类资料相比显示出种族和地域性差异,由此建立起了辽宁地区已知HPA基因型的血小板捐献者资料库。  相似文献   

6.
目的对邢台地区血小板志愿捐献者进行人类血小板抗原(human platelet antigen,HPA)基因分型,建立已知HPA基因型的血小板捐献者资料库。方法采用聚合酶链反应-序列特异性引物(polymerase chain reaction-sequence specific primers,PCR-SSP)方法对随机抽取的150例血小板捐献者进行HPA-1~6和HPA-1 5抗原系统共14个抗原基因分型。结果邢台地区血小板志愿捐献者HPA的基因频率分别为:HPA-1a 0.953,1b 0.01 0;2a 0.9 53,2b 0.086;3a0.596,3b 0.394;4a 1.000,4b 0:5a 0.967,5b 0.01 5:6a 0.986,6b 0.010;15a 0.475,1 5b 0.506。结论邢台地区血小板志愿捐献者HPA-1~6和HPA-15抗原系统的基因频率符合Hardy-weinberg遗传定律,与其他地区和国家的同类资料相比显示出种族和地域性差异,应建立邢台地区已知HPA基因型的血小板捐献者资料库。  相似文献   

7.
上海地区汉族人群人类血小板抗原基因的多态性研究   总被引:3,自引:0,他引:3  
目的研究上海地区汉族人群人类血小板抗原HPA-1~16系统基因多态性分布,为快速寻找更合适的血小板输注提供实验依据。方法采用PCR-SSP方法对137名上海地区汉族健康成年人进行HPA-1~16系统基因分型。结果HPA-1~6和HPA-15的基因频率分别为HPA-1a 0.9854,HPA-1b 0.0146,HPA-2a 0.9453,HPA-2b 0.0547,HPA-3a 0.5511.HPA-3b 0.4489,HPA-4a 0.9964,HPA-4b 0.0036,HPA-5a 0.9891,HPA-5 b0.0109,HPA-6a 0.9781,HPA-6b 0.0219,HPA-15a 0.5292,HPA-15b 0.4708;其余HPA系统只检测到a等位基因,其基因频率均为1.0000。经χ^2检验,符合Hardy-Weinberg遗传定律。结论HPA基因多态性分布存在明显的种族和地域性差异。HPA-1~6和HPA-15系统基因频率分布具有多态性,其中HPA-3和HPA-15系统杂合度最高,抗原分布不配合比例相对高,在临床配合性血小板输注中必须加以重视。HPA-3基因多态性分布更具有上海地区特点。  相似文献   

8.
目的:研究豫北地区汉族无偿献血人群血小板抗原(HPA-1-5、15)系统多态性,构建一定规模的血小板HPA基因资料库,为临床提供匹配的单采血小板。方法:随机采集豫北5地区500名汉族无偿献血者外周血,用PCR-SSP方法进行HPA-1-5、15系统基因分型。基因及基因型频率采用直接计数法检测,HPA基因频率的群体分布情况采用Hardy-weinberg平衡检验,与其他地区和种族比较采用χ~2检验。结果:豫北地区无偿献血人群HPA-1-5、15血型系统基因分别为1a0.985、1b0.015;2a0.924、2b0.076;3a0.469、3b0.531;4a1.000;5a1.000;15a0.532、15b0.468。HPA-1-5、15基因型频率分别为1aa0.970、1ab0.030;2aa0.848、2ab0.152;3aa0.222、3ab0.494、3bb0.284;4aa1.000;5aa1.000;15aa0.282、15ab0.500、15bb0.218。与中国汉族人群相比,豫北地区无偿献血人群HPA-3b、5a增高,差异有统计学意义(P﹤0.05);与中国少数民族人群相比,HPA-3b、5a也升高,但HPA-2a降低(P﹤0.05)。与国外不同种族人群相比,HPA-1a、2a、3a、5a均有不同程度的升高。结论:豫北地区无偿献血人群HPA基因分布符合Hardy-weinberg平衡定律。与其他地区和国家资料相比,HPA显示出种族和地域性差异。豫北地区无偿献血人群HPA-3、15最具多态性。这为研究同种免疫性血小板减少症和开展血小板同型输注的相关研究提供理论依据。  相似文献   

9.
目的研究汉族人群血小板特异性抗原HPA-1-16系统基因多态性区域分布特点,建立HPA基因型资料库。方法采用PCR-SSP技术对102名潍坊地区汉族无血缘关系的志愿捐献血小板者作HPA-1-16系统基因分型,计算基因型频率、基因频率。结果从HPA-1、5、6系统分别检出1例a/b基因型,HPA-4系统检出2例a/b基因型,检出HPA-2a/2b 11例(基因频率占0.1078);HPA-3、15系统表现出较高的杂合度,其中3a/3a、3a/3b、3b/3b基因型频率分别为0.3039、0.5392、0.1569,15a/15a、15a/15b、15b/15b基因型频率分别为0.3529、0.4608、0.1863;HPA- 7-14、16系统只检出a基因,呈单特异性。结论潍坊地区汉族人HPA-2、3、15系统具有多态性,是HPA配合性输注关注重点。在随机输血中供受者HPA-2系统不配合的机会为9.68%、HPA-3系统不配合的机会为36.95%、HPA-15系统不配合的机会为36.8%。  相似文献   

10.
本研究的目的是分析人类血小板抗原(human platelet antigen,HPA)基因多态性,根据分布频率来判断HPA抗原不配合比率以及抗体产生的机会,确定有临床意义的血小板抗原系统,并建立邯郸地区血小板基因频率数据库和供者库.采用SSP-PCR方法对邯郸地区148名随机献血者进行HPA1-16抗原32个等位基因的检测分析,并与不同人群的分布频率进行比较.结果表明:每个样本均检测到HPA-1a、2a、4a-14a、16a基因;HPA-4a、7a-14a、16a呈现单态性,未检测出相应的等位基因HPA-b;对于HPA-1、-2、-5、-6主要以a/a纯合子为多,a/a基因型频率分别是0.9595、0.8108、0.9865、0.9797,没有b/b纯合子出现.在HPA1-16中,具有最高杂合度的是HPA-15,基因型HPA15a/15a、HPA15a/15b、HPA15b/15b频率分别是0.2230、0.5270、0.2500;HPA-3在其次,基因型HPA3a/3a、HPA3a/3b、HPA3b/3b频率分别是0.3851、0.5135、0.1014.经x2检验,结果符合Hardy-Weinberg遗传平衡定律.邯郸地区随机献血者HPA1-5系统基因频率与石家庄地区相似(P>0.05);与我国台湾人群进行HPA1-13、HPA-15的比较,HPA-1、-2、-6具有明显的不同(P<0.05),其它相似(P>0.05);与韩国人群进行HPA1-8的比较,除HPA-3具有明显不同外(P<0.05),其余均相似(P>0.05);与美国黑人进行HPA1-5的比较,HPA-1、-2、-5具有明显的差异(P<0.05);与英国人进行HPA1-11的比较,HPA-1、-5具有明显的不同(P<0.05).结论:北方地区中国人群HPA-2、-3、5、-15系统具有多态性,且HPA抗原分布不配合比率较高,这必然造成免疫暴露的机会增加,提示在临床上可能具有重要的免疫学意义.同时,在此次研究数据的基础上建立了邯郸地区血小板基因频率数据库和血小板已知型供者库.  相似文献   

11.
Genotyping of the human platelet alloantigens (HPA) is useful for the diagnosis and therapy of the patients with alloimmune thrombocytopenic syndromes, such as post-transfusion refractoriness to platelets, post-transfusion thrombocytopenic purpura and foetomaternal alloimmune thrombocytopenia. We have developed, optimized and validated a new method for simultaneous genotyping of HPAs - HPA-1, HPA-2, HPA-3 and HPA-5 - by using the real-time polymerase chain reaction (PCR) based on TaqMan technology. Its performances were compared to those of the standard PCR-sequence-specific primers (SSP) method by testing 120 DNA samples. Several discrepancies between the two methods have been observed, especially in the HPA-3 genotyping. Evidently, the PCR-SSP method produced several false positive results due to its technical drawbacks. Based on our comparison, we believe that the new real-time TaqMan PCR assay for the HPA-1, HPA-2, HPA-3 and HPA-5 genotyping is faster, more reliable and reproducible, compared to the standard PCR-SSP.  相似文献   

12.
青岛地区汉族人群HPA-1—5,15多态性分布研究   总被引:11,自引:6,他引:5  
目的研究青岛地区汉族人群人类血小板抗原(HPA)1-5,15抗原分布多态性。方法采用PCR-SSP方法对青岛地区918名无血缘关系固定血小板无偿捐献者进行HPA1-5及HPA-15系统的基因分型.结果各被检系统等位基因频率分别是1a=0.9940,1b=0.0060,2a=0.9319,2b=0.0681,3a=0.5822,3b=0.4178,4a=0.9897,4b=0.0104,5a=0.9804,5b=0.0196,15a=0.4913,15b=0.5087;HPA基因频率分布与国内资料比较,HPA-1与北方人群(河南),HPA-2与南方人群(四川)差异有统计学意义;与台湾人群HPA-2,-4,与日本人群HPA-2,-3,-5,与美国黑人HPA-1,-2,-5,与白人HPA-1,-4,-5,-15分别有统计学显著性差异。结论青岛地区汉族人群HPA分布具有本地人群特点。本组HPA数据分布符合Hardy-Weinberg平衡定律,可以作为北方汉族人群HPA基因分布频率数据库和青岛本地化血小板供者HPA资料库。  相似文献   

13.
目的了解上海地区单采血小板献血人群HPA-1~5、15多态性分布,分析评估新的分型技术。方法利用TaqMan PCR技术对500份上海地区单采血小板供者标本进行HPA-1~5、15抗原系统等位基因分型,并随机抽取100份标本使用PCR-SSP技术进行比对。结果 HPA各等位基因频率分别为HPA-1a:0.999,HPA-1b:0.001,HPA-2a:0.953,HPA-2b:0.047,HPA-3a:0.582,HPA-3b:0.418,HPA-4a:0.999,HPA-4b:0.001,HPA-5a:0.988,HPA-5b:0.012,HPA-15a:0.524,HPA-15b:0.476;有1份标本HPA-5等位基因与SSP检测结果产生差异。结论上海地区HPA各等位基因频率与国内各地区人群分布无明显差异,与中国汉族人群HPA分布情况基本吻合,实验数据经验证符合Hardy-Weinberg平衡定律,实验结果准确可靠;HPA-5差异经测序验证判断可能由PCR-SSP非特异扩增所致;TaqMan技术在HPA抗原系统分型应用中特异性高,反应时间短,具有良好的应用前景,是现有技术方法的一种重要补充。  相似文献   

14.
目的 了解广州汉族献血人群人类血小板抗原(HPA)-21多态性.方法 设计HPA-21的序列特异性引物(SSP),采用SSP法对200名广州汉族无偿血小板献血者作HPA-21基因分型,并采用DNA测序验证.结果 200名献血者中,198例为HPA-21a/-21a纯合子,2例为HPA-21a/-21b杂合子,并经DNA测序证实.广州汉族献血人群HPA-21a和HPA-21b的等位基因频率分别为99.50%和0.50%,与Hardy-Weinberg平衡吻合;在随机输血中HPA-21抗原不配合的机会为0.99%.结论 所获得的广州汉族献血人群HPA-21基因频率数据,有助于建立本地血小板供者库和开展血小板同型输注.  相似文献   

15.
16.
BACKGROUND: Single-amino-acid substitution Leu33Pro in the beta3-integrin is responsible for the formation of the human platelet antigen (HPA)-1. Alloimmunization against HPA-1a (beta3-Leu33) is the most frequent cause of neonatal alloimmune thrombocytopenia and posttransfusion purpura. STUDY DESIGN AND METHODS: While HPA-1 genotyping a large cohort of patients with thromboembolic disease with a thermal cycler (LightCycler), one patient was identified with a unique HPA-1a melting curve. RESULTS: Sequence analysis revealed a C-to-G transversion at nucleotide 175 in the beta3-integrin (ITGB3) gene that alters the Leu33 codon to Val33. Further genotyping of healthy blood donors (n = 2950) identified one nonrelated Pro33Val33-positive individual. To examine whether the presence of Val33 affected the binding pattern of HPA-1 alloantibodies, transfectants were generated expressing recombinant beta3-Leu33 or beta3-Val33. Interestingly, differences in the reactivity of anti-HPA-1a were observed, with some HPA-1a alloantibodies showing diminished reactivity with beta3-Val33 compared to beta3-Leu33 and others reacting equally with both types. Similar findings were observed with recombinant human HPA-1a antibodies, with one of the three not binding to beta3-Val33. CONCLUSIONS: Our results demonstrate that the naturally occurring Leu33Val mutation in the beta3-integrin can disrupt some HPA-1a epitopes. These findings provide evidence for a heterogeneous humoral response against HPA-1a that may have potential clinical implications for alloimmune thrombocytopenia disorders.  相似文献   

17.
Non-invasive fetal HPA-1a typing is a valuable tool to identify the pregnancies at risk of fetal and neonatal alloimmune thrombocytopenia (FNAIT). At present, prenatal determination of the fetus HPA-1a type is performed for diagnostic purposes in pregnancies of HPA-1 alloimmunized women with history of a previous fetus or child with FNAIT. Different approaches have been used to determine the fetal HPA-1a genotype from cell-free fetal DNA (cffDNA) in the mother’s plasma, mainly based on real-time PCR. Due to the single nucleotide polymorphism (SNP) between the HPA-1a and HPA-1b allelic sequences, a robust and accurate detection of the fetal genotype is challenging, and the sensitivity of most assays is still limited early in pregnancy. Nowadays, the availability of technologies such as next generation sequencing (NGS) or digital PCR offers unprecedented possibilities of analyzing cell-free DNA (cfDNA)-amplified sequences with very high coverage and high sensitivity. In addition, other interesting approaches using variant sequence enrichment strategies have been recently described. In particular, coamplification at lower denaturation temperature PCR (COLD-PCR) offers a simple and sensitive strategy for noninvasive fetal HPA-1 typing. These novel approaches are explained in more detail in this review. Despite no population-based FNAIT screening programs have so far been implemented, the perspectives in terms of treatment and prevention are changing and less costly high-throughput maternal HPA-1a typing methods have been developed. Altogether, this may lead to the implementation of fetal HPA-1a typing with a broader scope in the future, playing a critical role within FNAIT screening programs.  相似文献   

18.
SUMMARY. We report a male patient who developed post-transfusion purpura. Thrombocytopenia did not recur when intravenous IgG was given before the transfusion of platelet-depleted washed red-cell concentrates.  相似文献   

19.
SUMMARY. The human platelet alloantigens HPA-1, -2, -3, -5 and -6b in the Finnish population were determined using allele specific restriction analysis (PCR-ASRA) for HPA-1, -2, -3 and -5 and monoclonal antibody immobilized platelet antigen (MAIPA) assay for HPA-1, -3a, -5b and -6b. No discrepancies were observed between the results obtained with the PCR-method and those obtained serologically. The gene frequencies obtained from 200 unrelated Finns were 0.86 and 0.14 for HPA-1a and -1b, 0.91 and 0.09 for HPA-2a and -2b, 0.59 and 0.41 for HPA.3a and -3b and 0.95 and 0.05 for HPA-5a and -5b. The frequency of the HPA-5b allele (10%) is lower in Finns than in Central- or South-European populations (20–30%). The HPA-1, -2 and -3 frequencies did not deviate from those observed in other European populations. The rare HPA-6b antigen was observed in three of 127 individuals from south-eastern Finland (2.4%), which suggests that the frequency of this allele in Finland is higher than previously thought.  相似文献   

20.
The human granulocyte antigen HNA-2a is expressed on a subpopulation of granulocytes in 97% of Caucasians. Alloimmunization against this antigen can cause alloimmune neonatal neutropenia, transfusion-related acute lung injury and immune neutropenia after bone marrow transplantation. Immunochemical studies demonstrated that NB1 allogenic determinants reside on a GPI-linked glycoprotein with apparent molecular weight of 56–64 kDa. To elucidate the primary structure of NB1 protein, we isolated NB1 protein using NB1-specific mab 7D8 by affinity chromatography. Identity of the purified glycoprotein was confirmed by immunoblot using a panel of seven human and two monoclonal NB1-specific antibodies. Determination of molecular mass using MALDI yielded a 50·6 kDa glycoprotein. After removal of N-linked carbohydrates, the molecular mass was reduced to 43·1 kDa. N-terminal amino acid sequencing by Edman degradation yielded a 20 aa-peptide. Rapid amplification of cDNA ends derived from an HPA-2a (+) individual using primers based on N-terminal amino acid sequence resulted in a 1614-bp cDNA, consisting of a 27-bp 5' untranslated region, a 1311-bp open reading frame and a 276-bp 3' untranslated region. The 1311 bp open reading frame encoded for 437 amino acids, of which the first 21 were found to form the signalpeptide. The identity of this transkript was confirmed by the expression of full lengh NB1 cDNA (bp 22–1394) in COS-7 cells.  相似文献   

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