Establishment and application of a reverse dot blot assay for 13 mutations of hearing-loss genes in primary hospitals in China

Abstract

Abstract Background Hearing loss is a common sensorineural dysfunction with a high incidence in China. Although genetic factors are important causes of hearing loss, hearing-related gene detection has not been widely adopted in China. Objective Establishing a rapid and efficient method to simultaneously detect hotspot hearing loss gene mutations. Methods A reverse dot blot assay combined with a flow-through hybridization technique was developed for the simultaneous detection of 13 hotspot mutations of 4 hearing loss–related genes including GJB2, GJB3, SLC26A4, and the mitochondrial gene MT-RNR1. This method involved PCR amplification systems and a hybridization platform. Results The technique can detect 13 hotspot mutations of 4 hearing loss–related genes. And a total of 213 blood samples were used to evaluate the availability of this method. Discussion Our reverse dot blot assay was a simple, rapid, accurate, and cost-effective method to identify hotspot mutations of 4 hearing loss–related genes in a Chinese population.