Design and validation of a metabolic disorder resequencing microarray (BRUM1).
Résumé
Abstract The molecular genetic diagnosis of inherited metabolic disorders is challenging. The diseases are rare and most show locus heterogeneity. Hence testing of the genes associated with IMDs is time consuming and often not easily available. We report a resequencing array that allows the simultaneous resequencing of up to 92 genes associated with IMDs. To validate the array, DNA samples from 51 patients with 52 different known variants (including point variants, small insertion and deletions (indels)) in 7 genes (C14ORF133, GAA, NPC1, NPC2, VPS33B, WFS1, SLC19A2) were amplified by PCR and hybridised to the array. A further patient cohort with 48 different mutations in NPC1 were analysed blind. Out of 76 point variants, 73 were identified using automated software analysis followed by manual review. Ten insertion and deletion variants were detected in the extra tiling using mutation specific probes whilst 11 heterozygous deletions and 3 heterozygous insertions. In summary, we identified 96% (95%CI 89-99%) of point variants added to the array, but the pickup rate reduced to 83% (95%CI 75-89%) when insertions/deletions were included. Whilst the methodology has strengths and weaknesses, application of this technique could expedite diagnosis in most patients with multi-locus IMDs.
Origine : Fichiers produits par l'(les) auteur(s)
Loading...