A sensitive and specific diagnostic test for hearing loss using a microdroplet PCR-based approach and next generation sequencing

作者：Schrauwen Isabelle; Sommen Manou; Corneveaux Jason J; Reiman Rebecca A; Hackett Nicole J; Claes Charlotte; Claes Kathleen; Bitner Glindzicz Maria; Coucke Paul; Van Camp Guy^*; Huentelman Matthew J

来源：American Journal of Medical Genetics, Part A, 2013, 161A(1): 145-152.

DOI：10.1002/ajmg.a.35737

• hearing loss
• deafness
• microdroplet PCR
• diagnostics

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 1020% 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.

• 出版日期2013-1