Abstract: Objective To perform whole exome sequencing for a deaf couple and their fetus to provide genetic counseling for the family. Methods The clinical data, the samples of peripheral blood from the pregnant woman and her spouse，and amniotic fluid from the pregnant woman were collected. The genomie DNAs were extracted. The whole exome sequencing for the family was performed to find suspected pathogenic mutation sites which were verified by Sanger sequencing. Mutation sites that may lead to abnormal selective shear were analyzed by Minigene techniqueThe minigene technique was used to analyze the possible alternative splicing caused by the mutation that might induce variable splicing. Results The pregnant woman carried suspected compound pathogenic heterozygous mutations in transmembrane serine protease 3( TMPRSS3) gene, and her spouse carried pathogenic homozygous mutation c. 235delC in GJB2 gene. The fetus carried compound heterozygous mutations in TMPRSS3 gene which involved in a uncertainly significant c.323-8T>C inherited from his father and a possibly pathogenic c.325 C>T inherited from his mother. The detection of minigene technology found that c.323. -8T>C heterozygous mutation did not lead to obvious splicing change. Conclusion The newly discovered compound heterozygous mutations in TMPRSS3 gene and homozygous mutations in GJB2 gene may cause deafness in the pregnant woman and her spouse respectively. The compound heterozygous mutations in TMPRSS3 gene carried in the fetus does not cause deaf，thus the couple have only a slim chance for a deaf child.