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3.
Immunohematology ; 31(2): 81-90, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26495894

RESUMO

Classic methods to determine human red blood cell (RBC) antigens are based on serologic testing. Thanks to increased knowledge of the molecular basis associated with many blood group antigens, it is currently possible to predict their presence or absence on the red cell membrane. Several molecular techniques have been developed to detect the most important allelic variations attributable to single nucleotide polymorphisms. The human erythrocyte antigen (HEA) BeadChip™ system manufactured by BioArray Solutions (Immucor, Warren, NJ) is one of the commercial DNA array platforms currently available to predict HEAs by DNA analysis. This technology provides a useful tool to increase the inventory of antigen-negative RBC units and prevent immunization of patients who require chronic transfusion by providing compatible RBC units based on matching by DNA testing.


Assuntos
Antígenos de Grupos Sanguíneos/genética , Eritrócitos/metabolismo , Técnicas de Genotipagem/métodos , Polimorfismo de Nucleotídeo Único/genética , Alelos , Antígenos de Grupos Sanguíneos/imunologia , Tipagem e Reações Cruzadas Sanguíneas/instrumentação , Tipagem e Reações Cruzadas Sanguíneas/métodos , DNA/sangue , DNA/genética , Transfusão de Eritrócitos/métodos , Eritrócitos/imunologia , Genótipo , Técnicas de Genotipagem/instrumentação , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único/imunologia , Reprodutibilidade dos Testes
5.
Transfusion ; 48(3): 505-12, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18067501

RESUMO

BACKGROUND: Traditionally, blood group typing has been performed with serologic techniques, the classical method being the hemagglutination test. Serotyping, however, may present important limitations such as scarce availability of rare antisera, typing of recently transfused patients, and those with a positive direct antiglobulin test. Consequently, serologic tests are being complemented with molecular methods. The aim of this study was to develop a low-cost, high-throughput method for large-scale genotyping of red blood cells (RBCs). STUDY DESIGN AND METHODS: Single-nucleotide polymorphisms associated with some clinically important blood group antigens, as well as with certain rare blood antigens, were evaluated: Jk(a)/Jk(b), Fy(a)/Fy(b), S/s, K/k, Kp(a)/Kp(b), Js(a)/Js(b), Co(a)/Co(b), and Lu(a)/Lu(b). Polymerase chain reaction (PCR)-amplified targets were detected by direct hybridization to microspheres coupled to allele-specific oligonucleotides. Cutoff values for each genotype were established with phenotyped and/or genotyped samples. RESULTS: The method was validated with a blind panel of 92 blood donor samples. The results were fully concordant with those provided by hemagglutination assays and/or sequence-specific primer (SSP)-PCR. The method was subsequently evaluated with approximately 800 blood donor and patient samples. CONCLUSION: This study presents a flexible, quick, and economical method for complete genotyping of large donor cohorts for RBC alleles.


Assuntos
Antígenos de Grupos Sanguíneos/genética , Eritrócitos/metabolismo , Polimorfismo de Nucleotídeo Único , Alelos , Eritrócitos/citologia , Genótipo , Humanos , Hibridização de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase/métodos , Software
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