A reverse dot blot assay for the screening of twenty mutations in four genes associated with NSHL in a Chinese population

作者：Li, Siping; Peng, Qi; Liao, Shengyun; Li, Wenrui; Ma, Qiang; Lu, Xiaomei^*

来源：PLos One, 2017, 12(5): e0177196.

DOI：10.1371/journal.pone.0177196

• NONSYNDROMIC HEARING-LOSS
• MOLECULAR ETIOLOGY
• DEAFNESS
• IMPAIRMENT
• SLC26A4
• GJB2
• CHILDHOOD
• MONGOLIA

Background @@@ Congenital deafness is one of the most distressing disorders affecting humanity and exhibits a high incidence worldwide. Most cases of congenital deafness in the Chinese population are caused by defects in a limited number of genes. A convenient and reliable method for detecting common deafness-related gene mutations in the Chinese population is required. @@@ Methods @@@ We developed a PCR-reverse dot blot (RDB) assay for screening 20 hotspot mutations of GJB2, GJB3, SLC26A4, and MT-RNR1, which are common non-syndromic hearing loss (NSHL)-associated genes in the Chinese population. The PCR-RDB assay consists of multiplex PCR amplifications of 10 fragments in the target sequence of the four above-mentioned genes in wild-type and mutant genomic DNA samples followed by hybridization to a test strip containing allele-specific oligonucleotide probes. We applied our method to a set of 225 neonates with deafness gene mutations and 30 normal neonates. @@@ Results @@@ The test was validated through direct sequencing in a blinded study with 100% concordance. @@@ Conclusions @@@ The results demonstrated that our reverse dot blot assay is a reliable and effective genetic screening method for identifying carriers and individuals with NSHL among the Chinese population.

• 出版日期2017-5-15