目标序列捕获及平行测序在临床耳聋基因诊断中的应用

发布时间：2018-10-17 14:20

【摘要】：目的利用目标序列捕获(Targeted sequence capture)、生物编码(Barcode)及大规模平行测序(Massivelyparallel sequencing,MPS)技术,对遗传性耳聋患者进行分子病因学研究,探索低成本、多基因、多样本的耳聋基因检测在临床应用的可行性。方法利用Illumina测序平台对来自解放军总医院聋病分子诊断中心88例(来自61个家系)具有明确家族史且常见耳聋基因检测阴性的耳聋患者,以及8例阳性对照患者进行42种基因的目标序列捕获、Barcode和MPS,并对测序结果进行生物信息学分析。结果利用上述方法在88个个体中大于一人入组家系中发现了10个家系8个基因(TECTA、DFNA5、CDH23、USH1C、MYO7A、POU3F4、SLC26A4、EYA4)14个位点的致病性突变。准确验证了8个阳性对照。大于一人入组家系的致病基因检出率明显高于单人入组家系。结论高通量测序的发展为遗传性耳聋的诊断带来了巨大的机遇。目标序列捕获、Barcode、MPS的结合进一步提高了测序的准确性、降低了测序成本,同时生物信息学的进步,将促进低成本、多基因、多样本的临床耳聋基因检测成为可能。
[Abstract]:Objective to study the molecular etiology of hereditary deafness patients by using target sequence capture (Targeted sequence capture), biocoding (Barcode) and large-scale parallel sequencing (Massivelyparallel sequencing,MPS) to explore low cost and multiple genes. The feasibility of multisample deafness gene detection in clinical application. Methods 88 deafness patients from the molecular diagnosis center of deafness in PLA General Hospital (61 families) with a clear family history and negative detection of common deafness genes were studied by Illumina sequencing platform. The target sequences of 42 genes were captured, Barcode and MPS, were analyzed by bioinformatics. Results the pathogenicity mutations at 14 loci of 8 genes (TECTA,DFNA5,CDH23,USH1C,MYO7A,POU3F4,SLC26A4,EYA4) in 10 families were found in more than one of 88 individuals. Eight positive controls were accurately verified. The detection rate of pathogenicity genes was significantly higher in families with more than one person. Conclusion the development of high-throughput sequencing presents great opportunities for the diagnosis of hereditary deafness. The combination of target sequence acquisition and Barcode,MPS further improves the accuracy of sequencing and reduces the cost of sequencing. At the same time, advances in bioinformatics will promote the detection of low cost, multi-gene and multi-sample clinical deafness genes.
【作者单位】：解放军总医院耳鼻咽喉头颈外科;解放军总医院海南分院耳鼻咽喉头颈外科;美国Emory大学耳鼻咽喉科;解放军总医院第一附属医院耳鼻咽喉头颈外科;解放军二炮总医院耳鼻咽喉科;
【基金】：十二五国家科技支撑计划重点项目(No.2012BAI12B00/2012BAI12B01) 中国博士后基金及特别资助(No.20120481482&201104779) 北京市自然科学基金(No.7122172)
【分类号】：R764.43

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