A simple and reliable method for the detection of the 30delG mutation of the CX26 gene

Mutations in the CX26 gene (GJB2), encoding the gap-junction protein Connex in-26, have been shown to be the major cause of non-syndromic recessive dea fness. Among these mutations, the deletion of a guanine within the stretch of six G between nucleotide positions +30 and +35 of the CX26 cDNA (30delG) accounts for the majority of this kind of deafness. Molecular detection of the 30delG mutation is usually performed by direct sequencing analysis of PCR products or by SSCP. To detect this mutation we developed an easy and r eliable method, based on PCR, followed by a non-radioactive sandwich hybrid ization on microtiter plates. We tested 188 individuals recruited from the genetic counseling service for deaf people at the Pasteur Hospital and at t he Armand-Trousseau Children's Hospital, Paris, France between April 1997 a nd September 1998. Our screening method is simple, uses stable and safe rea gents, and employs inexpensive equipment. As such, it is suitable for wides pread use in genetic diagnosis. (C) 2000 Academic Press.