The specific mutaiton, c.544_545insA in the EYA4 was prioritised due to, 1) The mutaiton co-segregate with hearing loss in the family, 2) The mutation was verified by Sanger sequencing in all 15 members and the results are consistent with those of the NGS, 3) The specific mutation was analyzed in 1000 age-matched unrelated random controls with normal hearing and no carrier was identified. Here, EYA4 is linked to hearing loss disorder.