Mass-scale high-throughput multiplex polymerase chain reaction for human platelet antigen single-nucleotide polymorphisms screening of apheresis platelet donors
| Autor: | Gregory A. Denomme, Nadine Shehata, Barbara Hannach, Nancy Banning, John Freedman |
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| Rok vydání: | 2011 |
| Předmět: |
endocrine system
biology Donor selection business.industry Immunology Plateletpheresis Hematology medicine.disease Molecular biology Human platelet antigen Antigen Multiplex polymerase chain reaction Neonatal alloimmune thrombocytopenia biology.protein Immunology and Allergy Medicine Multiplex business Genotyping hormones hormone substitutes and hormone antagonists |
| Zdroj: | Transfusion. 51:2028-2033 |
| ISSN: | 0041-1132 |
| DOI: | 10.1111/j.1537-2995.2011.03082.x |
| Popis: | BACKGROUND: Treatment with human platelet antigen (HPA)-matched platelets (PLTs) is the optimal therapy for bleeding secondary to neonatal alloimmune thrombocytopenia. Recent advances in high-throughput DNA-based blood group and PLT antigen genotyping have made it possible to screen plateletpheresis donors for potential HPA-matched PLT transfusion. STUDY DESIGN AND METHODS: This prospective study evaluated genomic DNA from plateletpheresis donors for single-nucleotide polymorphisms (SNPs) associated with HPA-1, -2, -3, -4, -5, and -15 to determine whether high-throughput multiplex genomic DNA PCR and oligonucleotide extension technology can be used for mass-scale PLT antigen genotyping. Genotyping using SNP technology was confirmed using sequence-specific polymerase chain reaction (SSP-PCR). RESULTS: Of the 748 donors screened, 277 were found to be negative for antigens implicated in alloimmune thrombocytopenia. In addition, two donors were homozygous for HPA-1b/b and -2b/b, six donors for HPA-1b/b and -3b/b, one for HPA-2b/b and -3b/b, one for HPA-1b/b and -5b/b, 10 for HPA-1b/b and -15 b/b, four for HPA-5b/b and -15b/b, and one for HPA-2b/b and -15b/b. Retesting using SSP-PCR was conducted for 60 donors. Discrepant results occurred between SNP and SSP-PCR in less than 20% of samples for HPA-1b/1b/HPA-3b/3b, HPA-5b/5b, and HPA-15b/b. DISCUSSION: High-throughput multiplex PCR SNP and confirmatory molecular genotyping are useful for mass-scale screening of apheresis PLT donors to provide antigen-negative genotypes. Refinements to mass-scale multiplex analysis technology would reduce further the confirmatory testing needed. |
| Databáze: | OpenAIRE |
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