Simultaneous Genotyping of Human Platelet Antigens by Hot Start Sequence‐Specific Polymerase Chain Reaction with DNA Polymerase AmpliTaq Gold

Chen, Dongfeng; Pastucha, L.T.; Chen, Haiyan; Kadar, J; Stangel, W

doi:10.1046/j.1423-0410.1997.7230192.x

Cited by 26 publications

(8 citation statements)

References 14 publications

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“…To this end, we determined the phenotype and allele frequencies for those systems in 33 German patients with chronic refractory AITP, and compared them with the data obtained from 80 healthy German blood donors. Allele frequencies from the controls agreed with published data [14]. Further, the allele frequencies of the HPA–1, –3, and –5 systems in the chronic refractory AITP patients were similar to the allele frequencies in controls indicating lack of association between alleles of the HPA–1, –3, and –5 systems and the development of chronic refractory AITP.…”

Section: Discussionsupporting

confidence: 85%

Allele Frequencies of Human Platelet Antigen 1, 2, 3, and 5 Systems in Patients with Chronic Refractory Autoimmune Thrombocytopenia and in Normal Persons

et al. 1999

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Background and Objectives: Chronic refractory autoimmune thrombocytopenic purpura (AITP) is an autoimmune disorder due to autoantibodies against platelet glycoproteins (GP). Human platelet alloantigenic (HPA) systems are distributed to different platelet GPs. We carried out genotyping of diallelic HPA–1, –2, –3, and –5 systems to clarify potential associations between HPA alleles and the development of chronic refractory AITP. Patients and Methods: DNA was isolated from 33 unrelated German patients with chronic refractory AITP and from 80 randomly selected German blood donors to determine the phenotype and allele frequencies for the HPA–1, –2, –3, and –5 systems. Fragments carrying the polymorphic sequences corresponding to those alleles were amplified by the polymerase chain reaction and further characterized by restriction analysis. Results: Whereas HPA–1, –3, and –5 allele frequencies were identical in 33 patients with chronic refractory AITP and in controls, HPA–2 allele frequencies showed a statistically significant difference (p = 0.017). In our group of patients, the HPA–2a allele frequency was 100%, but HPA–2b was not seen. In contrast, the allele frequency of HPA–2a in the control group was 92% (n = 147), and in HPA–2b it was 8% (n = 13). Conclusion: This study suggests an association between the HPA–2a allele and chronic refractory AITP. The HPA–2a allele may be involved in the formation of an AITP–specific autoepitope.

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Section: Discussionsupporting

confidence: 85%

Allele Frequencies of Human Platelet Antigen 1, 2, 3, and 5 Systems in Patients with Chronic Refractory Autoimmune Thrombocytopenia and in Normal Persons

et al. 1999

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“…In the past, HPA typing has been performed mostly by serological methods. Because of severe limitations of these methods, they have been replaced with rapid molecular methods such as polymerase chain reaction‐sequence specific primers (PCR‐SSP) (10–13) or PCR with restriction fragment length polymorphism (PCR‐RFLP) (14).…”

Section: Sequences Of Specific Primers Used For Human Platelet Antigementioning

confidence: 99%

Gene frequencies of human platelet antigens in the Macedonian population

Pavkovic

Petlichkovski²,

Strezova³

et al. 2006

Tissue Antigens

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Human platelet antigen (HPA) systems consist of more than 12 bi-allelic antigen polymorphisms. Due to these polymorphisms, platelet-membrane glycoproteins can be recognized as alloantigens or autoantigens and can cause conditions such as fetomaternal alloimmune thrombocytopenia, post-transfusion refractoriness to platelets, and post-transfusion throbocytopenic purpura. The purpose of this study was to investigate the distribution of HPA-1, -2, -3, and -5 in Macedonian population by using the polymerase chain reaction and restriction fragment length polymorphism. The allele frequencies were 0.865 for HPA-1a, 0.135 for HPA-1b, 0.852 for HPA-2a, 0.148 for HPA-2b, 0.578 for HPA-3a, 0.422 for HPA-3b, 0.909 for HPA-5a, and 0.091 for HPA-5b. Results of our study were not significantly different from those reported in the other European studies. Our population displayed the highest frequency for HPA-2b allele (0.148) reported among European population.

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“…Two primers which amplify a region of the C‐reactive protein (CRP) gene were used as an internal control in each amplification mixture. Genotyping of HPA systems was performed according to a previously published protocol with slight modifications in order to perform simultaneous determination of all HPA alleles (Chen et al , 1997). Briefly, the PCR mixture (20 μL) contained 1 × 10 × PCR buffer II, 0·5 U Amply Taq Gold, 0·2 mmol L −1 of each dNTP, 1·5 mmol L −1 MgCl 2 , 0·5 μmol L −1 of each specific primer, 0·05 μmol L −1 of the internal control primer (to amplify CRP) and 50 ng genomic DNA.…”

Section: Methodsmentioning

confidence: 99%

Gene frequencies of platelet-specific antigens in Croatian population

Pavić¹,

Zadro

Herak

et al. 2010

Transfusion Medicine

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The human platelet antigens (HPA) are genetically defined polymorphisms expressed on platelet membrane glycoproteins. As platelet antigens are very important in several clinical situations and in population genetics, we used the polymerase chain reaction with sequence-specific primers (PCR-SSP) to investigate HPA-1, -2, -3 and -5 allele frequencies in the Croatian population. The HPA frequencies obtained in 219 Croatians were: 1a-0.854, 1b-0.146, 2a-0.890, 2b-0.110, 3a-0.575, 3b-0.425, 5a-0.895 and 5b-0.105. These data are similar to the frequencies reported in most European studies with some significant differences in HPA-2 when compared with the Dutch and German population, in HPA-3 when compared with the Swiss population and in HPA-5 when compared with the Finnish population. The three most common condensed HPA genotypes in the Croatian population were: HPA-1a/a, -2a/a, -3a/b, -5-a/a (0.283), HPA-1a/a, -2a/a, -3a/a, -5-a/a (0.137) and HPA-1a/b, -2a/a, -3a/b, -5-a/a (0.087). Data obtained in this study can be used for better understanding and treatment of immune-mediated platelet disorders in our population.

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Simultaneous Genotyping of Human Platelet Antigens by Hot Start Sequence‐Specific Polymerase Chain Reaction with DNA Polymerase AmpliTaq Gold

Abstract: Using our hot start PCR-SSP procedure with AmpliTaq Gold a simple, rapid and reproducible genotyping for HPA-1, 2, 3 and S systems could be achieved.

Cited by 26 publications

References 14 publications

Allele Frequencies of Human Platelet Antigen 1, 2, 3, and 5 Systems in Patients with Chronic Refractory Autoimmune Thrombocytopenia and in Normal Persons

Allele Frequencies of Human Platelet Antigen 1, 2, 3, and 5 Systems in Patients with Chronic Refractory Autoimmune Thrombocytopenia and in Normal Persons

Gene frequencies of human platelet antigens in the Macedonian population

Gene frequencies of platelet-specific antigens in Croatian population

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