Platelet alloantigen frequencies in Amazon Indians and Brazilian blood donors

The frequencies of human platelet-specific alloantigens (HPAs) vary between different ethnic groups, and genotyping using DNA techniques has been preferred over immunophenotyping methods for population studies. Using a polymerase chain reaction with allele-specific primers (PCR-ASP) method, we determined the allelic polymorphisms of five HPA systems among 174 unrelated individuals of two different Brazilian ethnic groups including Amazon Indians (n = 95) and blood donors (n = 79). Comparison of the calculated gene frequencies of the two alleles of HPA-1, -2, -3, -4 and -5 systems for Amazon Indians and Brazilian blood donors showed that gene frequencies obtained for the two alleles of HPA-1 (P<0.001), HPA-2 (P = 0.001) and HPA-5 (P<0.001) were significantly different between the two groups of individuals. All natives tested carried the HPA-2a and the HPA-5a alleles, but the HPA-1b and HPA-4b alleles are absent from the Indian population. It was also observed that all blood donors carried the HPA-1a, HPA-4a and HPA-5a alleles. In conclusion, the present data indicate differences in the frequency of the HPA systems between Amazon Indians and Brazilian subjects who present a high rate of racial admixture. While the frequencies of the HPA-1 and HPA-5 genes seen in Amazon Indians are similar to those reported for Oriental populations, the frequencies of the HPAs alleles in Brazilian blood donors are comparable to those reported for populations in North America and Europe.     

广州地区无偿献血者HPA-1—6,15基因分型及频率调查

目的研究人类血小板基因多态性,为人类群体遗传学研究及临床输血实践提供重要数据和依据。方法通过PCR-SSP方法对广州地区706名无偿献血者的HPA1—6,15系统进行基因分型,并统计其频率。结果706份样本中HPA-3和-15基因型的杂合程度最高,其a/a、a/b、b/b的频率分别为HPA-3:0.2918、0.4830、0.2252;HPA-15:0.2691、0.5170、0.2139,不配合率较高,均达到35%以上。HPA-1、-2、-4、-5系统均以a/a纯合子为主,a基因的频率范围为0.9583—0.9993,且均未发现b/b纯合子。1b、4b的基因频率很低,分别为0.0028和0.0007。本地区的HPA-1a与中国北方、英国白人、美国黑人有统计学差异(P<0.05)。HPA-2与中国南方、北方、美国黑人和日本人有统计学差异(P<0.05);HPA-5与英国白人和美国黑人存在统计学差异(P<0.05)。HPA-6频率分别为0.9575,0.0397,0.0028。结论本研究对广州地区HPA献血员的筛查可为建立HPA供者库和对探讨由HPA引起的免疫性疾病的预防和治疗提供相关数据和研究手段。     

Genotyping of Eight Human Platelet Antigen Systems in Serbian Blood Donors: Foundation for Platelet Apheresis Registry

IntroductionThe aim of this study was to investigate the allele and genotype frequencies of 8 human platelet antigen (HPA) systems among blood donors from the Blood Transfusion Institute of Serbia and to compare them with published studies. These data would be useful to establish the basis for a platelet apheresis donor registry.Material and MethodsSeventy-two unrelated male platelet apheresis/blood donors from Serbia were typed for 8 HPA systems (HPA-1 to HPA-6, HPA-9, and HPA-15) via the FluoGene method, based on polymerase chain reaction-sequence-specific amplification (PCR-SSP; PCR using sequence-specific primers) with fluorometric signal detection. Allele and genotype frequencies were estimated by direct counting and compared to the expected genotype frequencies according to the Hardy-Weinberg principle. The transfusion mismatch probability was calculated for every HPA specificity.ResultsThe allele frequencies were: HPA-1a, 0.868; HPA-1b, 0.132; HPA-2a, 0.917; HPA-2b, 0.083; HPA-3a, 0.611; HPA-3b, 0.389; HPA-5a, 0.903; HPA-5b, 0.097; HPA-9a, 0.993; HPA-9b, 0.007; HPA-15a, 0.472; and HPA-15b, 0.528. For HPA-4 and HPA-6 only allele a was detected.DiscussionThe HPA allele frequencies of European populations showed no significant differences in comparison with our results. Statistically significant differences were revealed in comparison with some populations of non-European origin. In the tested donors no HPA-2 bb genotype was detected, but we found 1 donor with the rare HPA-9b allele. The biggest transfusion mismatch probability in the Serbian population is for systems HPA-15 (37.4%) and HPA-3 (36.2%), which means that more than a third of random transfusions could cause mismatch in these systems. This study was enabled by the introduction of molecular HPA typing, and it provides initial results of the HPA allele and genotype frequencies in the population of blood donors in Serbia. They will be used to provide a compatible blood supply on demand for treating patients with alloimmune thrombocytopenic disorders. The successful implementation of PCR-SSP with fluorometric signal detection could be further complemented in the future by the introduction of high-throughput methods, which will largely depend on the available financial resources.     

献血者血小板1-16抗原基因遗传多态性分析及已知HPA型供者数据库的建立

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.     

长春地区无偿献血者HPA1-6,15基因分型及频率调查

目的调查北方汉族人群人类血小板抗原（HPA）的基因频率及分布规律。建立一支HPA-1a,-2a,-4a,-5a,-6a阴性及已知HPA抗原的献血者队伍。方法随机抽取172名长春地区固定无偿血小板捐献者静脉血3ml,枸橼酸钠抗凝;采用TIANGEN试剂盒提取DNA。基因定量仪测定DNA的浓度和纯度。采用PCR-SSP方法进行试验。最后进行统计学分析。结果各表现型的频率分别是：HPA-1aa 0.988,HPA-1ab 0.012,HPA-2aa 0.833,HPA-2ab 0.159,HPA-2bb 0.008,HPA-3aa 0.355,HPA-3ab 0.482,HPA-3bb 0.163,HPA-4aa 0.988,HPA-4ab 0.012,HPA-5aa 0.971,HPA-5ab0.029,HPA-6aa 0.977,HPA-6ab 0.023,HPA-15aa 0.286,HPA-15ab 0.498,HPA-15bb 0.216。未发现HPA 1bb、4bb、5bb、6bb。各基因型的频率分别是：HPA-1a 0.994,HPA-2a 0.913,HPA-3a 0.596,HPA-4a 0.994,HPA-5a 0.985,HPA-6a 0.988,HPA-15a 0.535。其中基因频率占99%以上的有HPA-1a-、4a。各基因型的不配合率分别是HPA-1a 0.011,HPA-2a0.147,HPA-3a 0.366,HPA-4a 0.011,HPA-5a 0.028,HPA-6a 0.023,HPA-15a 0.374。结论本研究有助于在长春地区建立已知HPA抗原的机采血小板捐献者队伍,可以提高血小板抗体的检出率,为同种免疫性血小板减少症患者提供HPA相合的血小板。     

Establishment of an HPA-1- to -16-typed platelet donor registry in China

In order to determine gene frequencies of human platelet antigen (HPA) and establish a panel of accredited HPA-1a, -2a, -4a, -5a and -6a-negative donors as well as an HPA-typed platelet donor registry, a total of 1000 Chinese donors of Han nationality (500 from north China and 500 from south China) were typed for HPA-1 through -16 using a DNA-based polymerase chain reaction with sequence-specific primers genotyping method. The gene frequencies of HPA-1b, -2b, -3b, -4b, -5b, -6bw, -10bw and -15b were 0.0060, 0.0485, 0.4055, 0.0045, 0.0140, 0.0135, 0.0005 and 0.4680, respectively. The HPA-7bw, -8bw, -9bw, -11bw, -12bw, -13bw, -14bw and -16bw alleles were not found. The HPA-2b and -5b homozygous donors were detected at low frequencies. The HPA mismatch probabilities potentially leading to alloimmunization in random platelet transfusion vary with a region from 0.1% to 37% depending on the distribution patterns of common and less common alleles in each system. This study provides a useful HPA-typed plateletpheresis donor registry in China and could improve platelet antibody detection and HPA-matched platelet transfusion in alloimmune thrombocytopenic patients.     

海南黎族人血小板1—17抗原基因与随机输血不配合率研究

目的调查海南岛黎族人群血小板抗原(HPA)-1—17等位基因多态性及其特点,评估其在随机输血中血小板输注无效的风险。方法采用PCR-SSP方法对海南岛180名黎族人群做HPA-1—17基因分型检测>计算各系统对偶抗原不配合率。结果在海南黎族人群的HPA-1—17系统中,呈多态性分布的等位基因及其频率为HPA-2a(0.9972)、2b(0.0028),HPA-3a(0.4889)、3b(0.5111),HPA-5a(0.9667)、5b(0.0333),HPA-6a(0.9972)、6b(0.0028),HPA-15a(0.4250)、15b(0.5750);其余HPA-1、4、7—14、16、17等位基因均呈单线性分布。在HPA-3、15等位基因中出现bb纯合子基因型,频率分别为0.2834和0.3667;其余系统均未见bb纯合子基因型。随机输血中,海南岛黎族人群HPA不配合的发生率依次为:HPA-3(37.49%)、-15(36.93%)和-5(6.23%)。结论揭示了海南岛黎族人HPA-1—17基因型和等位基因频率的分布及特点;立足人群的随机血小板输注,只需检测供、受者HPA-2,-3、-5和-15基因相合,就可基本达到血小板匹配性输注。     

Human platelet-specific antigens frequencies in the Argentinean population

summary .  The frequencies of several human platelet antigens (HPAs) vary between different populations and are a major determinant for the prevalence of HPA alloimmunization and its clinical associated entities. The aim of this study was to characterize the allele frequencies of seven HPA systems in two different ethnic groups from the Argentinean city of Rosario, the major population and a minority Amerindian group recently arrived from the north of the country, the Tobas. A total of 192 healthy unrelated individuals from blood donors and hospital staff from the Hospital Italiano Garibaldi and 27 unrelated Toba Amerindians were genotyped for HPA-1, -2, -3, -4, -5, -6 and -15 systems by polymerase chain reaction with sequence specific primers (PCR-SSP). The present data showed that the distribution of the HPA alleles among Argentineans from Rosario is quite similar to that reported among Europeans. The frequencies seen in Tobas, although limited by the small number of aboriginal samples studied, are similar to those reported for other Amerindians populations. Statistically significant differences were found for the genotype distribution of HPA-1, -3, -5 and -15 between both groups, indicating important differences in the potential risk of HPA alloimmunization associated to transfusion and pregnancy. The study of these polymorphisms represents the first step in the elucidation of pathological conditions that are underdiagnosed in our population. It allowed us to establish a panel of characterized blood donors necessary for the serological work out and as a source for compatible platelets transfusion.     

张家口地区机采血小板捐献者HPA分型研究

目的:分析河北省张家口地区血小板捐献者血小板特异性抗原HPA1-5,15系统的基因多态性。方法:采集血小板捐献者静脉血并提取DNA,采用聚合酶链-序列特异性引物(PCR-SSP)法进行血小板特异性抗原分型,计算基因频率、基因型频率并与国内外其他地区进行比较是否存在差异及是否表现地区特异性。结果:HPA-1、HPA-2、HPA-4系统中基因表达均为纯合子aa,未出现表达纯合子bb的存在者,其中HPA-1和HPA-4系统各发现杂合子ab表达1例(1%),HPA-2系统发现杂合子ab表达14例(14%);HPA-5系统基因表达主要为纯合子aa(98%),极少数表达纯合子bb(2%);HPA-3和HPA-15系统基因表达杂合程度较高,HPA-3系统基因表达aa、ab、bb表达比例分别为46%、40%、14%,HPA-15系统基因表达aa、ab、bb表达比例分别为21%、64%、15%。结论:张家口地区血小板特异性抗原HPA1-5,15系统基因频率具有本地区特点;HPA-3和HPA-15系统基因表达杂合程度较高,引发同种免疫及血小板输注无效的可能性较大,因此应引起注意。     

人类血小板抗原1—16基因与血小板输注无效风险研究

目的探讨HPA-1—16基因多态性分布与血小板输注无效相关性。方法应用PCR-SSP法对上海地区268名汉族人群行HPA-1—16基因检测;应用ELISA法对49名反复输血的恶性血液病患者行血小板抗-HLA-Ⅰ与抗-HPA筛查试验。结果上海地区汉族人群HPA-1—16系统中,HPA-1—6,15系统等位基因频率1a=0.9889,1b=0.0111,2a=0.8881,2b=0.1119,3a=0.5989,3b=0.4011,4a=0.9963,4b=0.0037,5a=0.9907,5b=0.0093,6a=0.9832,6b=0.0168,15a=0.6418,15b=0.3582,均呈多态性分布;其余HPA-7—14,16系统等位基因均呈单线性分布。HPA-1,2,4—6,15系统主要以aa纯合子基因型频率分别为0.9776,0.7799,0.9925,0.9813,0.9664,0.4328。在HPA-2、3、15系统中出现bb纯合子基因型,其频率均为0.0037,0.1530,0.1492外,其余系统均未出现bb纯合子基因型。另外,在HPA-1—6,15系统中出现ab杂合子基因型,其频率分别为0.0224,0.2164,0.4963,0.0075,0.0187,0.0336,0.4180,以HPA-3杂合度最高,其次次序为HPA-15、HPA-2。在随机输血中,HPA不合发生率以HPA-3为最高(0.3650),其次分别为HPA-15(0.3541)、HPA-2(0.1790)。49名反复输血的恶性血液病患者中,有61.22%(30名)输血后相继出现抗-HLA-Ⅰ,而始终未检出抗-HPA。结论上海地区汉族人群血小板输注无效的主要原因是抗-HLA-Ⅰ所致;只需检测供者与受(患)者的HPA-2、-3、-15基因相合,就可基本达到血小板匹配性输注。     

深圳地区汉族人类血小板抗原1-6系统基因多态性分析

目的 研究人类血小板抗原基因多态性,为人类学研究及临床输血实践提供依据。方法 采用PCR-SSP方法对深圳地区222名汉族随机献血者HPA1-6系统进行基因分型研究,对其基因及基因型频率进行统计,并与HPA在不同人群中的分布进行对比分析。结果 在6个HPA系统中,HPA-3基因型的杂合程度最高,HPA-3a/3a、HPA-3a/3b、HPA-3b/3b的频率分别为0.265 8,0.518 0,0.216 2;其余5个HPA系统均以a/a纯合子为主,a基因的频率范围为0.997 7～0.955 0,且均未发现b/b纯合子。1b、4b的基因频率很低,分别为0.009 O和0.002 3。结论 深圳汉族人群HPA1-6系统的基因频率与中国台湾人及中国香港人均很相似(P>0.05)。HPA-1、HPA-5与美国黑人、白人及荷兰人差异显著(P<0.05);HPA-2、HPA-3与日本人、韩国人、美国黑人、白人差异显著(P<0.05);HPA-4与日本人差异显著(P<0.05)。在未来的临床实践中要警惕HPA-2,3,5,6系统同种抗体导致的同种免疫血小板减少综合征的可能。     

青岛地区汉族人群HPA-1—5,15多态性分布研究

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。     

新疆汉族人群血小板抗原1—5、15系统基因多态性分析

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。     

应用PCR—SSP法分析中国人血小板抗原基因型频率

本研究建立了在同一PCR反应条件下同时检测人血小板抗原（human platelet antigen,HPA）系统HPA-1到HPA-5的PCR-SSP检测法。应用该方法分析了110例健康献血员的HPA-1到HPA-5的基因型,并以此为依据推算了中国人HPA-1到HPA-5各亚型的基因频率。结果表明HPA-1a和HPA-1b的基因频率分别为0.91和0.09,HPA-2a和HPA-2的基因频率分别为0.86和0.14,HPA-3a和HPA-3b的基因频率分别为0.60和0.40,HPA-4a和HPA-4b的基因频率分别为0.92和0.08,HPA-5a和HPA-5b的基因频率分别为0.85和0.15。结论：基因组DNA的血小板抗原PCR-SSP分型法切实可行,可广泛应用于临床血小板抗原的分型。     

岳阳地区无偿献血者HPA1-6,15基因分型及频率调查

目的 了解岳阳地区汉族人群血小板抗原1-6,15多态性分布及基因频率.方法 采用PCR-SSP方法对岳阳地区204名无血缘关系固定血小板无偿捐献者进行HPA1-6及HPA-15系统的基因分型.结果 岳阳地区无偿献血者HPA1-6,15基因分型未发现HPA-1bb、-2bb、-4bb、-5bb.其中基因频率占98％以上的有HPA-1aa、-4aa、-5aa,本研究首次发现HPA-6bb,说明HPA-6具有多态性,杂合程度较低.结论 岳阳地区汉族人群HPA分布具有本地人群特点,调查数据分布符合Hardy-Weinberg平衡定律,HPA-3和15系统具有多态性,在同种免疫性血小板随机输注时易产生同种免疫反应.     

The use of genotyping to predict the phenotypes of human platelet antigens 1 through 5 and of neutrophil antigens in Taiwan

BACKGROUND: The human platelet antigen (HPA) 1 through 5 and the human neutrophil antigen (HNA-1) systems are relevant to immune-related thrombocytopenia and neutropenia. The alloantigen distribution profiles in the population will aid in estimating the risk of alloimmunization. STUDY DESIGN AND METHODS: Genotyping of the genes that control the expression of the HPA-1 through -5 and HNA-1 systems in Taiwanese (n = 326) and Taiwan's indigenous peoples (n = 608) was performed by PCR with the sequence-specific primer (PCR-SSP) method. RESULTS: In the HPA system, HPA-1b and HPA-4b were absent among Taiwan's indigenous tribes and detected among other Taiwanese only with frequencies of <0.2 percent and <0.5 percent, respectively. The GP1BA*2 (HPA-2b) and GP1A*2 (HPA-5b) allele frequencies range from 1 percent to 7 percent and 0.4 percent to 3.5 percent among the two ethnic groups, respectively. GP2B*1 (HPA-3a) and GP2B*2 (HPA-3b) showed similar allele frequencies. In the HNA-1 system, the FCGR3B*1 (HNA-1a) allele frequency was about twice that of FCGR3B*2 (HNA-1b) in Taiwanese and also in most of the indigenous tribes. Three FCGR3B (HNA-1) null persons were found in one indigenous tribe (Ami tribe), for an FCGR3B null frequency of 19.8 percent. However, no FCGR3B*3 (HNA-1c) allele was detected in Taiwan. CONCLUSION: The frequencies of HPA-1b, -2b, and -5b in the Taiwanese population were much lower than those among whites. In Taiwan, all of the HNA-1 null found was due to the deletion of the FCGR3B gene, and this deletion may be widely distributed in the Ami tribe.     

中国汉族人群HPA分型单采血小板供者库的建立模式与库容

本研究探讨建立中国汉族人群HPA分型单采血小板供者库的模式和合适的库容水平。研究方法为合并分析公开发表的中国16个省份的汉族血小板献血者人群人类血小板抗原（human platelet antigen,HPA）分布多态性群体调查数据。对HPA多态性群体调查数据进行Hardy—Weinberg平衡检验。结果表明,HPA．1、-4、石、-10未检出bb纯合子,HPA-7-9,11-14、-16未检出b基因。HPA1-16系统相互独立存在于血小板表面。中国汉族人群中发现了648种HPA组合,其中42种频率高于0．001的组合的累积频率达0．9763,频率最高（0．2012）的组合为：HPA-（7-8-9-11-12-13-14—16）aa-（1-4-5-6-10）aa-2aa-3ab-15ab。HPA对偶抗原不配合概率高于0．1的系统有HPA．15、-3和-2,对偶抗原不配合概率介于0．01—0．1的系统有HPA-1、-5和-6。中国汉族人群供受者间随机输血HPA1—16系统对偶抗原全匹配概率为0．3195。根据库容（N）随频率（F）变化的曲线,推导得到在P=95％时的回归方程LogN=-0．4394x Ln（F）＋0．4324。使用该公式椎测综合频率（HPA和AB0的频率乘积）为0．005的库容至少需要为576．07人。结论：中国汉族人群HPA分型单采血小板供者库应采用单库容600名供者的区域多中心联合建库模式,它既能解决当地主要的HPA—15、3和2系统不配合造成的血小板输注无效（platelet transfusion refractoriness,PTR）,还可以在遗传背景相近的区域有效地扩大建库规模以解决HPA低频抗原的不配合问题。在评估HpA分型血小板供者库容时不仅要考虑HPA的频率还同时要考虑ABO血型分布频率对库容的影响。     

2458名中国汉族人类HPA-1～6、15基因多态性的研究

目的分析中国汉族人群HPA-1～6、15系统的基因多态性,研究中国南、北方汉族人群HPA-1～6、15的基因分布。方法采用Luminex结合序列特异性寡核苷酸探针(flow-sequence specific oligonucleotide probes,FLOW-SSO)方法对2 458名深圳巿机采血小板无偿捐献者(其中南方人群1 554人,北方人群904人)进行HPA-1～6,15系统基因分型。采用聚合酶链反应-序列特异性引物(PCR-sequence specific primer,PCR-SSP)方法对有疑问的标本及罕见抗原(如HPA-1b/1b、2b/2b、5b/5b等)做进一步确认。结果中国汉族人群中HPA-1～6、15基因频率分别为,HPA-1a 0.991 7,HPA-1b 0.008 3,HPA-2a 0.955 2,HPA-2b 0.044 8,HPA-3a 0.558 6,HPA-3b 0.441 4,HPA-4a0.998 0,HPA-4b 0.002 0,HPA-5a 0.986 2,HPA-5b 0.013 8,HPA-6a 0.985 6,HPA-6b 0.014 4,HPA-15a 0.545 2,HPA-15b 0.454 8。南、北方汉族人群HPA-1～6、15比较,在HPA-1和HPA-3系统(χ2=15.032 0、5.418 8,P0.05);基因频率分布差异有统计学意义。经χ2检验,符合Hardy-Weinbery遗传定律。结论中国汉族人群中HPA-3和HPA-15杂合程度最高,在中国南北方汉族人群中HPA-1和HPA-3系统基因多态性分布存在明显的地域差异。为了给免疫性血小板减少症患者提供相合血小板输注,在中国建立血小板供者资料库是必要的。     

使用Taqman PCR技术建立人类血小板抗原-1～5、15分型体系

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。     

血小板谱抗原的建立及其在鉴定血小板特异性抗体中的应用

目的 建立血小板谱抗原,鉴定引起血小板输注无效和新生儿血小板减少性紫癜的血小板特异性抗体,为血小板血型研究和临床治疗提供依据。方法根据中国人群人类血小板同种抗原(HPA)-1-HPA-16等位基因频率分布资料,利用聚合酶链反应-序列特异引物(PCR-SSP)技术对O型血小板供者进行HPA-1-HPA-6、HPA-15分型,筛选合适的供者,组成血小板谱抗原。通过建立的血小板谱抗原,利用简易致敏红细胞血小板血清学技术(SEPSA)鉴定同种免疫反应产生的血小板抗体的特异性。结果从O型血小板供者中筛选出11名供者,建立了血小板特异性抗体鉴定谱抗原。其可鉴定HPA-1-HPA-6,HPA-15抗体的特异性。在所筛检1 120份样本中,有3例患者检出HPA抗体,其中HPA-4b(Penb)抗体1例,HPA-15a(Govb)抗体2例。结论通过血小板谱抗原鉴定血小板抗体的特异性,对提高临床输注血小板的安全性和有效性,以及预防新生儿血小板减少性紫癜有积极的意义.