RESUMEN
Pharmacogenetic testing is increasingly available from clinical laboratories. However, only a limited number of quality control and other reference materials are currently available to support clinical testing. To address this need, the Centers for Disease Control and Prevention-based Genetic Testing Reference Material Coordination Program, in collaboration with members of the pharmacogenetic testing community and the Coriell Cell Repositories, has characterized 137 genomic DNA samples for 28 genes commonly genotyped by pharmacogenetic testing assays (CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A5, CYP4F2, DPYD, GSTM1, GSTP1, GSTT1, NAT1, NAT2, SLC15A2, SLC22A2, SLCO1B1, SLCO2B1, TPMT, UGT1A1, UGT2B7, UGT2B15, UGT2B17, and VKORC1). One hundred thirty-seven Coriell cell lines were selected based on ethnic diversity and partial genotype characterization from earlier testing. DNA samples were coded and distributed to volunteer testing laboratories for targeted genotyping using a number of commercially available and laboratory developed tests. Through consensus verification, we confirmed the presence of at least 108 variant pharmacogenetic alleles. These samples are also being characterized by other pharmacogenetic assays, including next-generation sequencing, which will be reported separately. Genotyping results were consistent among laboratories, with most differences in allele assignments attributed to assay design and variability in reported allele nomenclature, particularly for CYP2D6, UGT1A1, and VKORC1. These publicly available samples will help ensure the accuracy of pharmacogenetic testing.
Asunto(s)
Proteínas Portadoras/genética , Sistema Enzimático del Citocromo P-450/genética , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/genética , Glucuronosiltransferasa/genética , Glutatión Transferasa/genética , Farmacogenética/métodos , Secuencia de Bases , Línea Celular , Pruebas Genéticas , Genotipo , Técnicas de Genotipaje , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Polimorfismo de Nucleótido Simple/genética , Análisis de Secuencia de ADN , Estados UnidosRESUMEN
The use of fetal cells enriched from maternal peripheral blood for genetic analysis would be a significant advancement for prenatal diagnosis. For over two decades, researchers have employed different techniques (including density gradients, red blood cell lysis, fluorescence-activated cell sorter and magnetic cell separation) to enrich fetal cells that exist in low levels in maternal blood. Despite extensive attempts by various groups and companies, reliable isolation of a sufficient amount of fetal nucleated cells remains a monumental challenge. AVIVA Biosciences Corp. has been developing a technology that allows significant enrichment and recovery of rare fetal cells from maternal peripheral blood. This review will characterize fetal cell types present in maternal blood, technologies used to enrich for these cells and the issues involved with rare fetal cell separation.
Asunto(s)
Células Sanguíneas/citología , Feto/citología , Separación Celular/métodos , Centrifugación por Gradiente de Densidad/métodos , Eritroblastos/citología , Femenino , Predicción , Humanos , Intercambio Materno-Fetal , Embarazo , Diagnóstico Prenatal/métodosRESUMEN
AutoGenomics, Inc. (Carlsbad, CA, USA) have developed a multiplex microarray assay for genotyping both VKORC1 and CYP2C9 using the INFINITI(™) Analyzer. Multiple alleles in each DNA sample are analyzed by polymerase chain reaction amplification, followed by detection primer extension using the INFINITI Analyzer. The INFINITI Analyzer performs single-nucleotide polymorphism (SNP) analysis using universal oligonucleotides immobilized on the biochip. To genotype broader ethnic groups, genomic DNA from whole blood was tested for nine SNPs for VKORC1 and six for CYP2C9 genotypes. Information related to all 15 SNPs is needed to determine dosing of population of diverse ethnic origin. The INFINITI system provides genotyping information for same day dosing of warfarin.