PDF(Pubmed)
Abstract：
Deafness is a common sensory disorder. In China, approximately 70% of hereditary deafness originates from four common deafness-causing genes: GJB2, SLC26A4, GJB3, and MT-RNR1. A single-tube rapid detection method based on 2D-PCR technology was established for nine mutation sites in the aforementioned genes, and Sanger sequencing was used to verify its reliability and accuracy. The frequency of hotspot mutations in deafness genes was analysed in 116 deaf students. 2D-PCR identified 27 genotypes of nine loci according to the melting curve of the FAM, HEX, and Alexa568 fluorescence channels. Of the 116 deaf patients, 12.9% (15/116) carried SLC26A4 mutations, including c.919-2A > G and c.2168A > G (allele frequencies, 7.3% and 2.2%, respectively). The positivity rate (29.3%; 34/116) was highest for GJB2 (allele frequency, 15.9% for c.235delC, 6.0% for c.299_300delAT, and 2.6% for c.176-191del16). Sanger sequencing confirmed the consistency of results between the detection methods based on 2D-PCR and DNA sequencing. Common pathogenic mutations in patients with non-syndromic deafness in Changzhou were concentrated in GJB2 (c.235delC, c.299_300delAT, and c.176-191del16) and SLC26A4 (c.919-2A > G and c.2168 A > G). 2D-PCR is an effective method for accurately and rapidly identifying deafness-related genotypes using a single-tube reaction, and is superior to DNA sequencing, which has a high cost and long cycle.
摘要：
耳聋是一种常见的感觉障碍。在中国,大约70%的遗传性耳聋源于四个常见的致聋基因:GJB2、SLC26A4、GJB3和MT-RNR1。针对上述基因中的9个突变位点,建立了基于2D-PCR技术的单管快速检测方法,并采用Sanger测序法验证其可靠性和准确性。分析了116名聋哑学生的耳聋基因热点突变的频率。根据FAM的熔解曲线，2D-PCR鉴定了9个基因座的27个基因型，HEX,和Alexa568荧光通道.116名聋哑患者中,12.9%（15/116）携带SLC26A4突变，包括c.919-2A>G和c.2168A>G(等位基因频率，7.3%和2.2%，分别)。GJB2阳性率(29.3%;34/116)最高(等位基因频率,c.235delC的15.9%，6.0%，c.299_300delAT，和2.6%的c.176-191del16）。Sanger测序证实了基于2D-PCR和DNA测序的检测方法之间结果的一致性。常州市非综合征性耳聋患者常见致病突变集中在GJB2(c.235delC,c.299_300delAT,和c.176-191del16）和SLC26A4（c.919-2A>G和c.2168A>G）。2D-PCR是一种利用单管反应准确快速鉴定耳聋相关基因型的有效方法,优于DNA测序，成本高、周期长。
