title: A sensitive and specific diagnostic test for hearing loss using a microdroplet PCR-based approach and next generation sequencing.
abstract: Implementing DNA diagnostics in clinical practice for extremely heterogeneous diseases such as hearing loss is challenging, especially when attempting to reach high sensitivity and specificity in a cost-effective fashion. Next generation sequencing has enabled the development of such a test, but the most commonly used genomic target enrichment methods such as hybridization-based capture suffer from restrictions. In this study, we have adopted a new flexible approach using microdroplet PCR-based technology for target enrichment, in combination with massive parallel sequencing to develop a DNA diagnostic test for autosomal recessive hereditary hearing loss. This approach enabled us to identify the genetic basis of hearing loss in 9 of 24 patients, a success rate of 37.5%. Our method also proved to have high sensitivity and specificity. Currently, routine molecular genetic diagnostic testing for deafness is in most cases only performed for the GJB2 gene and a positive result is typically only obtained in 10-20% of deaf children. Individuals with mutations in GJB2 had already been excluded in our selected set of 24 patients. Therefore, we anticipate that our deafness test may lead to a genetic diagnosis in roughly 50% of unscreened autosomal recessive deafness cases. We propose that this diagnostic testing approach represents a significant improvement in clinical practice as a standard diagnostic tool for children with hearing loss.
authors.affiliation: Department of Medical Genetics, University of Antwerp, Antwerp, Belgium.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Schrauwen
authors.forename: Manou
authors.initials: M
authors.lastname: Sommen
authors.forename: Jason J
authors.initials: JJ
authors.lastname: Corneveaux
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Reiman
authors.forename: Nicole J
authors.initials: NJ
authors.lastname: Hackett
authors.forename: Charlotte
authors.initials: C
authors.lastname: Claes
authors.forename: Kathleen
authors.initials: K
authors.lastname: Claes
authors.forename: Maria
authors.initials: M
authors.lastname: Bitner-Glindzicz
authors.forename: Paul
authors.initials: P
authors.lastname: Coucke
authors.forename: Guy
authors.initials: G
authors.lastname: Van Camp
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Huentelman
mesh_terms: D000072259:Connexin 26; D017630:Connexins / Q000235:genetics; D003638:Deafness / Q000175:diagnosis* / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D017630:Connexins; C000607042:GJB2 protein, human; D000072259:Connexin 26