RHD and RHCE genotyping by next-generation sequencing is an effective strategy to identify molecular variants within sickle cell disease patients
| Autor: | Francisco C. A. Gomes, Lucas A M Franco, José Eduardo Krieger, Sandra Fátima Menosi Gualandro, Karen Z. Chinoca, Ingrid Helena Ribeiro, Leonardo Varuzza, Ester Cerdeira Sabino, Marcia Regina Dezan, José Eduardo Levi, Carla Luana Dinardo, Alexandre C. Pereira, V.B. Oliveira, Alfredo Mendrone-Junior, Vanderson Rocha, Roberto Ribeiro, Juliana B. Vieira |
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| Rok vydání: | 2017 |
| Předmět: |
Genotype
Locus (genetics) Disease Anemia Sickle Cell 030204 cardiovascular system & hematology Biology DNA sequencing Serology 03 medical and health sciences 0302 clinical medicine Isoantibodies Genetic variation Humans Blood Transfusion Genetic Testing Allele Molecular Biology Genotyping Alleles Genetics Rh-Hr Blood-Group System Disease Management Genetic Variation High-Throughput Nucleotide Sequencing Reproducibility of Results Cell Biology Hematology Phenotype Molecular Medicine SEQUENCIAMENTO GENÉTICO 030215 immunology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| ISSN: | 1096-0961 |
| Popis: | Background The complexity of Rh genetic variation among sickle cell disease (SCD) patients is high. Conventional molecular assays cannot identify all genetic variants already described for the RH locus as well as foresee novel alleles. Sequencing RHD and RHCE is indicated to broaden the search for Rh genetic variants. Aims To standardize the Next Generation Sequencing (NGS) strategy to assertively identify Rh genetic variants among SCD patients with serologic suspicion of Rh variants and evaluate if it can improve the transfusion support. Methods Thirty-five SCD patients with unexplained Rh antibodies were enrolled. A NGS-based strategy was developed to genotype RHD and RHCE using gene-specific primers. Genotype and serological data were compared. Results Data obtained from the NGS-based assay were gene-specific. Ten and 25 variant RHD and RHCE alleles were identified, respectively. Among all cases of unexplained Rh antibodies, 62% had been inaccurately classified by serological analysis and, of these, 73.1% were considered as relevant, as were associated with increased risk of hemolytic reactions and shortage of units suitable for transfusion. Conclusion The NGS assay designed to genotype RH coding regions was effective and accurate in identifying variants. The proposed strategy clarified the Rh phenotype of most patients, improving transfusion support. |
| Databáze: | OpenAIRE |
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