Candidate Gene Screening For Autosomal Dominant Auditory Neuropathy In A Chinese Pedigree

Objective:To investigate if the OTOF gene,DFNB59 gene and MPZ gene contribute to the non-syndromic hearing loss of a Chinese pedigree with dominantly inherited auditory neuropathy(AN)using candidate approach.Methods:Nine live members in four generations of a kernel pedigree with dominantly inherited auditory neuropathy,3 sporadic AN patients and 3 normal-hearing individuals were selected for this study.Genomic DNA was isolated from the peripheral leukocytes of the subjects using the Puregene DNA Isolation Kits.The whole coding sequence of OTOF gene,DFNB59 gene,and MPZ gene of one family patient were PCR amplified using specific primers. Each fragment was purified and subsequently analyzed by direct sequencing in an Applied Biosystems 3730 automated DNA sequencer.The resultant sequence data were compared with the standard sequence to identify deafness-associated mutations.Other DNA samples were then screened for these mutations by PCR amplification and sequence analysis as described above.Results:PCR amplifications were successfully conducted in all the subjects.Comparison of the resultant OTOF sequence in one family patient with the standard sequence identified 10 new nucleotide variants which do not lead to amino acid change.These mutations were also detectable in other family individuals,3 sporadic AN patients and 3 normal-hearing controls.For the DFNB59 gene and MPZ gene,however,we failed to detect the presence either of the mutations that have been reported,or any other deafness-associated mutations by sequence analysis.Conclusions:Sequence analysis of OTOF gene,DFNB59 gene and MPZ gene reveals no meaningful mutations.These genes seem not contribute to the pathogenesis of this Chinese AN family,which suggesting new gene(s) involvement.