Design and application of a 23-gene panel by next-generation sequencing for inherited coagulation bleeding disorders

Autor: Maria Luisa Lozano, Luis Javier García-Frade, Ramón García-Sanz, M. del Rey, Rosa Fisac, M. J. Cebeira, Rafael Ramos, José María Bastida, José Ramón González-Porras, C. Aguilera, M. E. Sarasquete, J.M. Hernández-Rivas, Marcos González-Díaz, Emilia Pardal, M. E. Fontecha, B. Pérez, Susana Riesco, Rocío Benito, J. M. Martin-Antorán, Nuria Bermejo, M. C. Mendoza, Carlos Aguilar, María Paz Martínez, M. R. Cardesa, L. A. Silvestre
Rok vydání: 2016
Předmět:
Adult
Male
0301 basic medicine
Haemophilia
Adolescent
Genotype
Genetic counseling
Mutation
Missense
Prenatal diagnosis
030204 cardiovascular system & hematology
Biology
DNA sequencing
Inherited rare bleeding disorders
Frameshift mutation
Fibrinogen disorders
Young Adult
03 medical and health sciences
symbols.namesake
Blood Coagulation Disorders
Inherited
0302 clinical medicine
medicine
Humans
Genetic Testing
Child
Frameshift Mutation
Genotyping
Genetic Association Studies
Genetics (clinical)
Genetic testing
Genetics
Sanger sequencing
medicine.diagnostic_test
High-Throughput Nucleotide Sequencing
Infant
DNA
Sequence Analysis
DNA
Hematology
General Medicine
Middle Aged
Inherited coagulation factor deficiencies
030104 developmental biology
Child
Preschool
Next-generation sequencing
symbols
Female
Molecular diagnosis
Gene Deletion
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
ISSN: 1351-8216
Popis: [Introduction]: Molecular testing of Inherited bleeding coagulation disorders (IBCDs) not only offers confirmation of diagnosis but also aids in genetic counselling, prenatal diagnosis and in certain cases genotype–phenotype correlations are important for predicting the clinical course of the disease and to allow tailor-made follow-up of individuals. Until recently, genotyping has been mainly performed by Sanger sequencing, a technique known to be time consuming and expensive. Currently, next-generation sequencing (NGS) offers a new potential approach that enables the simultaneous investigation of multiple genes at manageable cost. [Aim]: The aim of this study was to design and to analyse the applicability of a 23-gene NGS panel in the molecular diagnosis of patients with IBCDs. [Methods]: A custom target enrichment library was designed to capture 31 genes known to be associated with IBCDs. Probes were generated for 296 targets to cover 86.3 kb regions (all exons and flanking regions) of these genes. Twenty patients with an IBCDs phenotype were studied using NGS technology. [Results]: In all patients, our NGS approach detected causative mutations. Twenty-one pathogenic variants were found; while most of them were missense (18), three deletions were also identified. Six novel mutations affecting F8, FGA, F11, F10 and VWF genes, and 15 previously reported variants were detected. NGS and Sanger sequencing were 100% concordant. Conclusion: Our results demonstrate that this approach could be an accurate, reproducible and reliable tool in the rapid genetic diagnosis of IBCDs.
Databáze: OpenAIRE
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