OBJECTIVETo investigate the polymorphisms of platelet membrane glycoprotein genes related to human platelet alloantigen (HPA)-1 to 17w.

METHODSThe DNA segments of platelet membrane glycoprotein genes related to HPA-1 to 17w were amplified using author's designed primers. The amplification products were purified and directly sequenced to identify the HPA genotype and glycoprotein gene polymorphisms.

RESULTSThirteen new single nucleotide polymorphisms (SNPs) and a micro-satellite sequence were found in the glycoprotein genes from the 112 random samples, in which two SNPs (1333G/A and 1960G/A) in ITGB3 gene result in two amino acid change (V419M and E628K) on glycoprotein GPIIIa.

CONCLUSIONNew variants in platelet membrane glycoprotein genes were identified, which may lead to structure change of platelet membrane glycoprotein and affect the accuracy of partial HPA genotyping method.

Antigens, Human Platelet ; genetics ; Humans ; Isoantigens ; genetics ; Platelet Membrane Glycoproteins ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVE: To study the polymorphism of human platelet antigen (HPA) system 10 among ethnic Han Chinese from Shandong, China so as to supplement the data of platelet donor bank in the region. METHODS: Peripheral blood samples of platelet donors from the region were genotyped for HPA-10 alleles by PCR-sequence specific primer (PCR-SSP) and direct sequencing. RESULTS: Among 1401 donors, a rare heterozygote carrier of HPA-10w (a+b+) was identified, which gave an allelic frequency of approximately 0.035%. CONCLUSION The detection of rare HPA-10bw antigen allele among ethnic Han Chinese from Shandong is useful for the diagnosis and prevention of neonatal alloimmune thrombocytopenia and post-transfusion purpura in the region.
Alleles ; Antigens, Human Platelet/genetics* ; Asians/genetics* ; Gene Frequency ; Genotype ; Humans ; Infant, Newborn ; Polymorphism, Genetic

Human platelet alloantigens (HPA) are specific antigens carried by platelet glycoproteins, which genes showing single nucleotide polymorphism. HPA can induce alloantibodies bringing about alloimmune response. They play important roles in post-transfusion refractoriness to platelets, post-transfusion thrombocytopenic purpura, fetomaternal alloimmune thrombocytopenia, and graft-versus-host disease. Because of their side effects in clinical blood-transfusion, there were a great deal of studies on HPA during last few decades. This review focuses on the nomenclature of HPA, the polymorphisms of platelet glycoproteins, HPA typing of the serological and molecular technology, as well as the mechanism of alloimmunization to HPA and correlated diseases.
Antigens, Human Platelet ; classification ; immunology ; Humans ; Isoantibodies ; immunology ; Platelet Membrane Glycoproteins ; genetics ; Polymorphism, Single Nucleotide ; Transfusion Reaction

OBJECTIVE: To study the Polymorphism of the human platelet antigen(HPA) gene 1-17 and human leukocyte antigen(HLA) gene-A and B locus in Shandong Han population. METHODS: A total of 962 samples from routine voluntary platelet donors were genotyped for HPA1-17 system and HLA-A site, B by PCR-SSP and PCR-SSOP respectively．Gene frequencies were calculated by counting. HPA1-17 and HLA genotype combinations were analyzed by Arelequin 3.5. RESULTS: The gene frequencies of HPA-la, -1b, HPA-2a, -2b, HPA-3a, -3b, HPA-4a, -4b, HPA-5a, -5b, HPA-6a, -6b, HPA-15a, -15b were 0.9918, 0.0082, 0.9419, 0.0592, 0.5841, 0.4174, 0.9969, 0.0031, 0.9892, 0.0108, 0.9835, 0.0175,0.5488 and 0.4512, respectively． The most common HPA genotype combination was HPA-(1, 2, 4, 5, 6, 7-14, 16, 17) aa-3ab-15ab (0.2048). Moreover, HLA-A*2(0.3094) and HLA-B*13(0.1513) showed the highest frequency in their respective locus. The most common HLA genotype combination was HLA-A*2-B*13(0.1397) . CONCLUSION Distributions of HPA and HLA show high polymorphism in Shandong Han population. The ethnic and territorial difference of HPA distribution is also confirmed. It is imperative to establish local genetic database of volunteer platelet donors.
Alleles ; Antigens, Human Platelet/genetics* ; Gene Frequency ; Genotype ; Humans ; Polymorphism, Genetic

OBJECTIVE: To analyze the polymorphism of the HPA1-5,15 system of the donors in Zhangjiakou area. METHODS: DNA was extracted from the blood samples of the donors, PCR- SSP method was used to divide HPA1-6, 15 genotype. The gene frequency and genotype frequency were calculated, compared with the difference and regiahal specificity of the populations in our country and foregiens was compared other populations. RESULTS: The gene expression in the HPA-1, HPA-2 and HPA-4 systems were all homozygous aa, and the donors who expressed homozygous bb was not exessed. Among them, one heterozygous ab expression was found in both HPA-1 and HPA-4 systems (1%), and 14 cases of heterozygous ab expression were found in HPA-2 system (14%). The gene expression in the HPA-5 system was mainly homozygous aa (98%), and a very few expressed homozygous bb (2%) was found. The degree of heterozygosity of gene expression in the HPA-3 and HPA-15 systems was relatively high. The proprotion of the expression of aa, ab and bb in the HPA-3 system was respectively 46%, 40% and 14%, the proprotion of the expression of aa, ab and bb in the HPA-15 system was respectively 21%, 64% and 15%. CONCLUSION The gene frequency of platelet-specific antigen HPA1-5,15 system in zhangjiakou region shows local characteristics. The heterozygosity degree of gene expression in the HPA-3 and HPA-15 systems are both high, suggesting that they are more likely to result in alloimmunization and ineffective platelet transfusion, which should be pays attention to.
Antigens, Human Platelet/genetics* ; Blood Donors ; Gene Frequency ; Genotype ; Humans ; Polymorphism, Genetic

The purpose of this study was to investigate the allele frequencies of the human platelet alloantigens 1-18 system (HPA-1-18) in Chinese Nanjing unrelated and healthy Han population, so as to provide the credible basis to screen compatible platelets for transfusing patients. The genotypes of 18 HPA systems were determined by polymerase chain reaction using sequence-specific primer (PCR-SSP) for 300 samples. The results showed that the gene frequencies obtained from 300 Nanjing unrelated population were 0.9183 and 0.0817 for HPA-2a and -2b, 0.6100 and 0.3900 for HPA-3a and -3b, 0.9733 and 0.0267 for HPA-5a and -5b, 0.9883 and 0.0117 for HPA-6a and -6b, 0.5250 and 0.4750 for HPA-15a and -15b. All the tested individuals were homozygotes for HPA-1a, -4a, -7a-14a and HPA -16a-18a. There was a good fit to Hardy-Weinberg equilibrium in each group. It is concluded that this study has confirmed the ethnic and regional difference of HPA, and HPA in Nanjing Han population has its own characteristics. The highest heterozygotes are HPA-3 and HPA-15, thus more attention to HPA effects on clinical platelet matched transfusion should be paid.
Antigens, Human Platelet ; genetics ; Asian Continental Ancestry Group ; genetics ; China ; Ethnic Groups ; genetics ; Genotype ; Heterozygote ; Homozygote ; Humans ; Polymorphism, Genetic

This study was aimed to investigate the detection and diagnosis of the neonatal alloimmune thrombocytopenia purpura (NAITP) caused by anti-HPA-5b antibody. The platelet count and clinical manifestation in the newborn were examined. The HPA-1-21bw genotypes of the newborn and her parents were detected by multiple-PCR and DNA sequencing. The HPA-specific antibody in the sera of newborn and her mother were detected and identified by flow cytometry (FCM) and monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The results indicated that the clinical manifestations of the newborn were lighter. The HPA genotyping showed that the genotype of the newborn was HPA-5ab, while that of her mother and father were HPA-5aa and HPA-5ab, respectively. The antibody against the platelet of newborn's father existed in the newborn's mother sera. The HPA antibody of the mother was identified as anti-HPA-5b. It is concluded that the newborn with neonatal alloimmune thrombocytopenia purpura was caused by the antibody against HPA-5b.
Antigens, Human Platelet ; genetics ; China ; Female ; Genotype ; Humans ; Infant, Newborn ; Purpura, Thrombocytopenic, Idiopathic ; diagnosis ; genetics ; Thrombocytopenia, Neonatal Alloimmune ; diagnosis ; genetics

This study was aimed to establish the reliable genotyping method of human platelet antigens of HPA-15 system by PCR-SSP and to use this assay in the further HPA genotyping of volunteer platelet donors. 3 sequence-specific primers recommended by the 11th Platelet Genotyping and Serology Workshop on behalf of International Society of Blood Transfusion (ISBT) were synthesized. The concentration of each primer pair, the concentration of Mg(2+) and the PCR conditions were adjusted to optimize the conditions so that HPA-15 system could be specific amplified. The accuracy and reliability of the developed assay was evaluated and confirmed by typing the coded DNA samples provided by the 11th Platelet Genotyping and Serology Workshop. As a parallel control, a total of 50 volunteer platelet donors in Shenzhen were genotyped by both our assay and the G&T commercial kit at HPA-15 system. 10 coded samples distributed by the 11th Platelet Genotyping and Serology Workshop were genotyped by established PCR-SSP method. The results showed that a concordance rate of 100% was observed between the results obtained by established PCR-SSP method and the results provided by ISBT report. The HPA gene frequencies observed in 50 randomly-selected platelet donors in Shenzhen were 0.5100 and 0.4900 for HPA-15a and HPA-15b respectively. In conclusion, PCR-SSP assay established in our study provides a simple, rapid and accurate method for HPA-15 system genotyping, which assay is suitable for routine clinical HPA genotyping and shows a broad prospect in its further applications.
Antigens, CD ; genetics ; immunology ; Antigens, Human Platelet ; genetics ; GPI-Linked Proteins ; Genotype ; Humans ; Isoantigens ; genetics ; immunology ; Neoplasm Proteins ; genetics ; immunology ; Polymerase Chain Reaction ; methods ; Polymorphism, Single-Stranded Conformational

This study was purposed to determine the mode and size of human platelet antigens (HPA) typed platelet apheresis donor bank. The published data of HPA distribution collected from Chinese Han population of 16 provinces were analyzed. The combined data were tested with the Hardy-Weinberg equilibrium. The results showed that the bb homozygote was not detected in HPA-1, -4, -6, -10, and b gene was not found in HPA-7-9, 11-14, -16. There were 648 combined HPA 1-16 genotypes in Chinese Han population, and the cumulative frequency of 42 combinations higher than 0.001 were 0.9763. The highest frequency (0.2012) in combination was HPA-(7-8-9-11-12-13-14-16) aa - (1-4-5-6-10) aa-2aa-3ab-15ab. The probability of HPA dual antigen mismatch in HPA-15, -3 and -2 was higher than the 0.1, and the probability in the HPA-1, -5, and -6 was between 0.01 - 0.1. The probability of full-match in HPA1-16 antigens was 0.3195 in Chinese Han population after the random blood transfusion. According to the curve drawn by donor number (N) versus frequency (F), the regression equation LogN = -0.4394 x Ln (F) +0.4324 was derived at P = 95%. If the derived frequency (product of HPA frequency and ABO frequency) is 0.005, then the N should be 576.07 at least in Chinese Han population. It is concluded that the mode of regional, multi-center database of HPA-typed platelet apheresis donor bank may be acceptable in Chinese Han population, and the suitable number of HPA-typed platelet donor in one bank may be 600. Therefore, the bank can be used to treat the platelet transfusion refractoriness (PTR) caused by HPA-15, 3 and 2 mismatch mainly, and can be expanded effectively in similar genetic background to deal with the low-frequency HPA antigens mismatch. The number of HPA-typed platelet apheresis donor influences not only on the frequency of HPA, but also on the frequency of ABO group.
Antigens, Human Platelet ; genetics ; immunology ; Asian Continental Ancestry Group ; genetics ; Blood Donors ; Gene Frequency ; Genotype ; Histocompatibility Testing ; Humans ; Platelet Transfusion ; Plateletpheresis

OBJECTIVETo study the polymorphism of human platelet alloantigens HPA-3 and HPA-9w in the Chinese Han population.

METHODSA total of 1000 unrelated Chinese Han blood donors from different provinces of China were genotyped for HPA-3 and HPA-9w using PCR-sequence specific primer assay.

RESULTSGene frequencies of 1000 Chinese Hans for HPA-3a and HPA-3b were 0.5935 and 0.4065 respectively, and all of them were HPA-9a positive. The distributions of HPA-3, HPA-9w of Chinese Hans which detected by chi-square criterion fit Hardy-Weinberg equilibrium. There were significant differences of the HPA-3 alleles gene frequency between Guangdong province and other five investigated provinces which included Shanxi, Heilongjiang, Zhejiang, Yunnan and Jiangsu. In comparison to other ethnic groups, no significant differences were observed in the distributions of HPA-3 except the Vietnamese and Australian.

CONCLUSIONThe results show that the chance of HPA-3 incompatibility were 0.3661 in random transfusion, and also provide a basis for researching on alloimmune thrombocytopenia and HPA-matched transfusion.

Alleles ; Antigens, Human Platelet ; genetics ; immunology ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; DNA ; genetics ; Ethnic Groups ; genetics ; Gene Frequency ; Genotype ; Histocompatibility ; genetics ; Humans ; Polymorphism, Genetic