Computational protocol: Genetic Screening of Mutations Associated with Fabry Disease in a Nationwide Cohort of Juvenile Idiopathic Arthritis Patients

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Protocol publication

[…] We aimed to calculate the frequency of FD in a population of JIA patients by GLA genotyping. Carriers of Fabry’s-associated mutations were further characterized.Children and young adults (mean age 17.7 years and 11 years of mean disease duration) from a JIA cohort of patients consecutively included from Paediatric Rheumatology centres in Portugal Mainland and Islands. All patients were diagnosed by an expert physician in the field and classified according to the International League of Associations for Rheumatology (ILAR) criteria (). Clinical and laboratorial information was recorded at regular time points, according to clinical practice, in the Portuguese nationwide register, a questionnaire with clinical information and a laboratorial evaluation of relevant data in the context of JIA were also performed in these patients. Blood samples collected have been stored at the IMM Biobank, Lisbon Academic Medical Centre.This study was submitted to the Local Ethics Committee, at Hospital de Santa Maria, Lisbon Academic Medical Centre. For being included in the study, parents and patients gave their informed written consent.Molecular genetic testing to identify GLA mutations was performed in the Centre of Histocompatibility in Coimbra. Direct sequencing was performed for samples from all patients in order to identify the precise mutation. Multiplex polymerase chain reactions with primers located in intron–exon boundaries were performed in order to allow sequencing of the complete coding region of GLA gene (seven exons). The GLA gene sequencing was performed in 3130 ABI sequencing platform (Applied Biosystems), and data were analyzed with SeqScape v3.0 and Geneious R9 software. Direct sequencing of the appropriate amplified fragment was a feasible strategy due to the development of rapid and sensitive automated sequencers. Control samples from healthy individuals were run in parallel with patients’ samples in order to confirm the consensus sequence for all the amplified exons. […]

Pipeline specifications

Software tools SeqScape, Geneious
Application Sanger sequencing
Organisms Homo sapiens
Diseases Fabry Disease, Arthritis, Juvenile, Rheumatic Diseases, Lysosomal Storage Diseases