Rapid genotyping of blood group antigens by multiplex polymerase chain reaction and DNA microarray hybridization
- PMID: 15847653
- DOI: 10.1111/j.1537-2995.2005.04319.x
Rapid genotyping of blood group antigens by multiplex polymerase chain reaction and DNA microarray hybridization
Abstract (V体育平台登录)
Background: In the Netherlands, 500,000 blood donors are active. Blood of all donors is currently typed serologically for ABO, the Rh phenotype, and K VSports手机版. Only a subset of donors is typed twice for a larger set of red cell (RBC) and/or platelet (PLT) antigens. To increase the direct availability of typed RBCs and PLTs, a high-throughput technique is being developed to genotype the whole donor cohort for all clinically relevant RBC and PLT antigens. .
Study design and methods: A multiplex polymerase chain reaction was developed to both amplify and fluorescently label 19 gene fragments of RBC and PLT antigens in one reaction. To test the setup of the genotyping method by microarray, a pilot study with human PLT antigen (HPA)-typed donor samples was performed. On each slide, 12 arrays are present containing 20 probes per PLT antigen system (28 for HPA-3) V体育安卓版. The allele-specific oligohybridization method was used to discriminate between two different alleles. .
Results: Two blinded panels encompassing 94 donors were genotyped for HPA-1 through -5 and -15; no discrepancies were found compared to their serologic typing (HPA-1, -2, -3, -4, and -5) and genotyping (HPA-15; TaqMan, Applied Biosystems) V体育ios版. .
Conclusion: This study shows that the HPA microarray provides a reliable and fast genotyping procedure. With further development an automated throughput for complete typing of large donor cohorts can be obtained VSports最新版本. .
Comment in
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Goodbye to agglutination and all that?Transfusion. 2005 May;45(5):652-3. doi: 10.1111/j.1537-2995.2005.05052.x. Transfusion. 2005. PMID: 15847650 No abstract available.
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