title: A new, easy, and rapid high-throughput detection method for the common GJB2 (CX26), 35delG mutation.
abstract: GJB2 (Gap Junction protein beta type 2; Connexin 26, CX26) is known for its contribution to nonsyndromic recessive deafness (NSRD). One particular mutation, 35delG, a deletion of one guanine from a stretch of six leading to a frame shift early in the gene, has a high prevalence in populations from European descent. 35delG testing therefore has become a standard test in genetic diagnostic laboratories. Most of the currently available methods for the detection of 35delG are relatively time consuming, and not suited for high-throughput diagnostic testing. Within this paper we present a real-time PCR genotyping assay based on melting curve analysis, requiring only a single preparation step before the actual analysis. The assay was optimized on a panel of 48 samples with known 35delG genotypes and subsequently tested using a large Belgian population (N = 460) with unknown 35delG status. For the latter set of samples, real-time PCR results were validated with SNAPShot, an assay used in our laboratory for diagnostic purposes. The real-time PCR genotyping method has proven to be highly reliable, rapid, cost-effective, and suitable for high-throughput screening. We believe that this genetic test for 35delG will find widespread applications in the DNA diagnostic field.
authors.affiliation: Department of Medical Genetics, University of Antwerp, Antwerp, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Van Eyken
authors.forename: G
authors.initials: G
authors.lastname: Van Camp
authors.forename: J J
authors.initials: JJ
authors.lastname: Hendrickx
authors.forename: K
authors.initials: K
authors.lastname: Demeester
authors.forename: A
authors.initials: A
authors.lastname: Vandevelde
authors.forename: J Ben
authors.initials: JB
authors.lastname: Azza
authors.forename: P
authors.initials: P
authors.lastname: Van de Heyning
authors.forename: L
authors.initials: L
authors.lastname: Van Laer
mesh_terms: D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D005820:Genetic Testing / Q000191:economics / Q000379:methods*; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D013696:Temperature
chemical_list: D017630:Connexins; C000607042:GJB2 protein, human; D000072259:Connexin 26