Rapid determination of platelet alloantigen genotypes by polymerase chain reaction using allele-specific primers. 1994

B Skogen, and D B Bellissimo, and M J Hessner, and S Santoso, and R H Aster, and P J Newman, and J G McFarland
Cancer Diagnostics Laboratory, Blood Center of Southeastern Wisconsin.

BACKGROUND The technique of allele-specific polymerase chain reaction (PCR) amplification has been adapted for DNA-based human platelet alloantigen typing. METHODS Sequence-specific primers were used to discriminate between the alleles encoding the six major human platelet alloantigens in a series of patients and normal blood donors. RESULTS This technique allows the direct determination of platelet antigen genotypes from genomic DNA after PCR amplification and agarose gel electrophoresis. It offers significant advantages over previously described techniques for alloantigen identification, as the additional analytical steps of restriction enzyme digestion or dot blot hybridization are not required. The results obtained with this technique correlated precisely with those derived from serologic typing and allele-specific oligonucleotide hybridization. CONCLUSIONS The use of allele-specific PCR for platelet alloantigen typing should facilitate the development of DNA-based typing in other regional blood centers and clinical laboratories.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D005838 Genotype The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS. Genogroup,Genogroups,Genotypes
D006579 Heterozygote An individual having different alleles at one or more loci regarding a specific character. Carriers, Genetic,Genetic Carriers,Carrier, Genetic,Genetic Carrier,Heterozygotes
D006720 Homozygote An individual in which both alleles at a given locus are identical. Homozygotes
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000483 Alleles Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product. Allelomorphs,Allele,Allelomorph
D000939 Epitopes Sites on an antigen that interact with specific antibodies. Antigenic Determinant,Antigenic Determinants,Antigenic Specificity,Epitope,Determinant, Antigenic,Determinants, Antigenic,Specificity, Antigenic
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain
D016824 Antigens, Human Platelet Human alloantigens expressed only on platelets, specifically on platelet membrane glycoproteins. These platelet-specific antigens are immunogenic and can result in pathological reactions to transfusion therapy. Antigens, Platelet-Specific,Human Platelet Antigen,Human Platelet Antigens,Platelet Alloantigen,Platelet Alloantigens,Platelet-Specific Antigen,Platelet-Specific Antigens,Alloantigen, Platelet,Alloantigens, Platelet,Antigen, Human Platelet,Antigen, Platelet-Specific,Antigens, Platelet Specific,Platelet Antigen, Human,Platelet Antigens, Human,Platelet Specific Antigen,Platelet Specific Antigens

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