Ji Yu-bin, Han Dong-yi, Wang Da-yong, Zhou Yu, Zhao Cui, Wang Hui, Lan Lan, Wang Qiu-ju
Department of otolaryngology-Head & Neck Surgery, Chinese PLA General Hospital, Beijing, China.
Zhonghua Yi Xue Za Zhi. 2009 Sep 29;89(36):2531-5.
To discuss how to determine the number of samples in epidemiological study about deafness genes and reveal the characteristics of GJB2, SLC26A4 and mitochondrial DNA A1555G mutations in deaf-mute patients in schools for deaf-mutes in Shandong Province.
A total of 485 subjects were collected from the different schools for deaf-mutes in Shandong province. Amplified target fragments included GJB2 coding sequence, mtDNA12SrRNA and exon 8, 10, 17, 19 of SLC26A4 gene. The amplicons of mtDNA 12S rRNA were subjected to restriction enzyme Alw26I. The amplicons of patients whose enzyme reaction highly indicating A1555G mutation, amplicons of GJB2 and those exons PCR products of SLC26A4 were directly sequenced.
The study revealed that 36.29% patients had two mutated alleles (homozygote & compound heterozygote) of GJB2 (24.12%) and SLC26A4 (6.60%) and mtDNA12SrRNA A1555G (5.57%). The 235delC and IVS7-2A > G were still the mutational hot spot in GJB2 and SLC26A4 respectively.
The method of determining the number of sample is very important in the epidemiological study. There were about 24 thousand deaf-mute patients who were caused by three sensitive deafness genes mutations in Shandong province. Screening the sensitive deafness genes in newborn is imminent.
探讨如何确定耳聋基因流行病学研究中的样本量，并揭示山东省聋哑学校聋哑患者中GJB2、SLC26A4和线粒体DNA A1555G突变的特征。
从山东省不同聋哑学校收集485名受试者。扩增的目标片段包括GJB2编码序列、线粒体DNA 12S rRNA以及SLC26A4基因的第8、10、17、19外显子。对线粒体DNA 12S rRNA的扩增产物进行Alw26I限制性酶切。对酶切反应高度提示A1555G突变的患者的扩增产物、GJB2的扩增产物以及SLC26A4的那些外显子PCR产物进行直接测序。
研究显示，36.29%的患者具有GJB2（24.12%）、SLC26A4（6.60%）和线粒体DNA 12S rRNA A1555G（5.57%）的两个突变等位基因（纯合子和复合杂合子）。235delC和IVS7-2A>G仍然分别是GJB2和SLC26A4的突变热点。
在流行病学研究中确定样本量的方法非常重要。山东省约有2.4万名聋哑患者是由三种敏感性耳聋基因突变引起的。对新生儿进行敏感性耳聋基因筛查迫在眉睫。
