摘要
目的探讨耳聋易感基因流行病学筛查过程中合理选择样本量的方法和重要性；揭示山东省聋哑学校耳聋患者线粒体DNA12sRNAA1555G突变、GJB2和SLC26A4基因突变特征，为耳聋基因诊断方法和筛查策略提供数据基础。方法通过统计学方法确定样本量；应用聚合酶链反应对山东省485例聋哑学校耳聋患者的线粒体12SrRNA基因、GJB2和SLC26A4外显子8，10，17，19基因编码区进行扩增；限制性内切酶A1w26I检测线粒体DNAA1555G突变。对酶切提示A1555G突变的病例、全部GJB2和SLC26A4基因扩增产物进行DNA测序。结果在485例患者中，27例携带A1555G突变（5．57％）；167例具有G_IB2已知致病突变（34．34％），其中致病突变频率为24．12％；60例携带SLC26A4已知致病突变（12．37％），致病突变率为6．60％；235delC和IVS7—2A〉G分别是GJB2和SLC26A4的突变热点；山东省聋哑患者中约有36．29％由这3种基因突变导致耳聋。结论样本量的合理选择在耳聋易感基因流行病学调查中至关重要；山东省有约2．4万聋哑患者是由这3种基因突变所导致耳聋；在新生儿中开展耳聋易感基因的筛查工作刻不容缓。
Objective 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. Methods 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 Alw261. The amplicons of patients whose enzyme reaction highly indicating A1555G mutation ,amplicons of GJB2 and those exons PCR products of SLC26Ad were directly sequenced. Results The study revealed that 36. 29% patients had two mutated alleles （ homozygote ＆ compound heterozygote） of GJB2 （ 24. 12% ） and SLC26A4 （ 6. 60% ） and mtDNAI2SrRNA AI555G （5.57%）. The 235delC and IVS7-2A 〉 G were still the mutational hot spot in GJB2 and SLC26A4 respectively. Conclusion 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.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第36期2531-2535,共5页
National Medical Journal of China
基金
基金项目：国家“863”高技术研究发展计划（2006AA022181）
国家自然科学基金（30830104,30572016,30672310,30771203）
国家“973”重点基础研究发展计划（2007CB507400）
国家“十一五”科技支撑计划（2006BA102800,2007BA118812）
高等学校全国优秀博士学位论文作者专项基金（200463）
北京市科技计划重大项目（D0906005040291）
北京市重大专项课题基金（7070002）
