[Application of next generation sequencing in 3 Waardenburg syndrome].

Objective:To test the gene sequence of 3 patients with Waardenburg syndrome（WS） using the next generation sequencing technology in order to explore the possible mechanism of molecular genetics. Methods:Medical histories of the family members were collected. Physical examination, audiological evaluation and CT examination were performed. Peripheral blood was collected and DNA was extracted. The exon region of 159 deafness genes, 6 mitochondrial genes and 3 miRNAs of the proband were tested by next generation sequencing. The mutation sites of the possible pathogenic genes were obtained, subsequently, Sanger sequencing verification was performed on the proband and family members. Results:The first proband had a heterozygous mutation in exon 7 of MITF gene（NM_000248）: c.641_643delGAA; The second proband had a heterozygous mutation in exon 10 of MITF gene（NM_001354605）: c.1177-1G>A; The third proband had a heterozygous mutation in exon 5 of PAX3 gene（NM_181457）: c.587_593delCCTCAGC; The parents of the three probands verified by Sanger sequencing that there was no variation at the corresponding sites, and the above mutations were spontaneous mutations. Conclusion:Next generation sequencing can more comprehensively analyze information of the carried status and genetic rules of the disease-associated gene in WS families, and provide guidance for family reproductives.目的：应用新一代测序技术对临床确诊为Waardenburg综合征（WS）的3例患儿进行基因检测，探讨可能的分子遗传学机制。 方法：对通过病史采集、全身及耳科查体、听力及影像学检查确诊为WS的3例患儿，采用新一代测序技术检测耳聋相关的159个基因的外显子区，6个线粒体基因，3个miRNA，得到可能的致病基因突变位点后，再对先证者的家属进行Sanger测序验证。 结果：第1例先证者MITF基因（NM_000248）7号外显子出现1个杂合突变：c.641_643delGAA；第2例先证者MITF基因（NM_001354605）10号外显子出现1个杂合突变：c.1177-1G>A；第3例先证者PAX3基因（NM_181457）5号外显子出现1个杂合突变：c.587_593delCCTCAGC；3例先证者父母通过Sanger测序验证相应位点均无变异，以上突变均为自发突变。 结论：新一代测序技术能够分析WS家庭致病基因的携带状况和遗传规律等信息，对家庭再生育进行指导。.