Open AccessGenetic Analysis through OtoSeq of Pakistani Families Segregating Prelingual Hearing Loss
Author(s) -
Shahzad Mohsin,
Sivakumaran Theru A.,
Qaiser Tanveer A.,
Schultz Julie M.,
Hussain Zawar,
Flanagan Megan,
Bhinder Munir A.,
Kissell Diane,
Greinwald John H.,
Khan Shaheen N.,
Friedman Thomas B.,
Zhang Kejian,
Riazuddin Saima,
Riazuddin Sheikh,
Ahmed Zubair M.
Publication year - 2013
Publication title -
otolaryngology–head and neck surgery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.232
H-Index - 121
eISSN - 1097-6817
pISSN - 0194-5998
DOI - 10.1177/0194599813493075
Subject(s) - sanger sequencing , genetics , hearing loss , biology , locus (genetics) , genotyping , genetic linkage , microsatellite , allele , concordance , population , genotype , dna sequencing , audiology , medicine , gene , environmental health
Objective To identify the genetic cause of prelingual sensorineural hearing loss in Pakistani families using a next‐generation sequencing (NGS)–based mutation screening test named OtoSeq. Study Design Prospective study. Setting Research laboratory. Subjects and Methods We used 3 fluorescently labeled short tandem repeat (STR) markers for each of the known autosomal recessive nonsyndromic (DFNB) and Usher syndrome (USH) locus to perform a linkage analysis of 243 multigenerational Pakistani families segregating prelingual hearing loss. After genotyping, we focused on 34 families with potential linkage to MYO7A, CDH23 , and SLC26A4 . We screened affected individuals from a subset of these families using the OtoSeq platform to identify underlying genetic variants. Sanger sequencing was performed to confirm and study the segregation of mutations in other family members. For novel mutations, normal hearing individuals from ethnically matched backgrounds were also tested. Results Hearing loss was found to co‐segregate with locus‐specific STR markers for MYO7A in 32 families, CDH23 in 1 family, and SLC26A4 in 1 family. Using the OtoSeq platform, a microdroplet PCR‐based enrichment followed by NGS, we identified mutations in 28 of the 34 families including 11 novel mutations. Sanger sequencing of these mutations showed 100% concordance with NGS data and co‐segregation of the mutant alleles with the hearing loss phenotype in the respective families. Conclusion Using NGS‐based platforms like OtoSeq in families segregating hearing loss will contribute to the identification of common and population‐specific mutations, early diagnosis, genetic counseling, and molecular epidemiology.