应用全外显子组测序技术筛查罕见石骨症致病基因

Application of whole exome sequencing technique for the pathogenic gene screening of rare osteopetrosis

目的筛查石骨症患者的致病基因,为疾病的基因诊断及预后提供参考。方法收集石骨症患者临床资料及外周血标本,提取DNA,构建全外显子测序文库,进行高通量测序并结合生物信息学技术筛查石骨症致病基因。结果对2例石骨症患者的全外显子进行了分析,平均测序深度分别为169.38X、231.06X,其中患者1携带罕见突变TCIRG1(c.1305+2T>C)、TCIRG1(c.2008C>T)和CLCN7(c.1116C>T);患者2携带罕见突变CLCN7(c.857G>A),生物信息学分析表明患者携带的罕见突变均对基因产物的结构和功能具有不同程度的影响。结论全外显子测序可一次性筛查已知石骨症致病突变,是石骨症致病突变筛查的有效工具,2例石骨症患者的临床病征可能与患者携带的TCIRG1和CLCN7突变密切相关。

Objective To screen the pathogenic gene of osteopetrosis to provide reference for its genetic di-agnosis and prognosis. Methods The clinical data and peripheral blood samples were collected from the pa-tients with osteopetrosis？ DNA was extracted, the whole exome sequencing library was built, then the high throughput detection was performed and the pathogenic gene was screened by combining with the bioinformat-ics technology. Results The whole exomes in 2 cases of osteopetrosis were analyzed？ the average sequencing depth of the two samples were 169. 38X and 231. 06X respectively in which the case 1 carried rare mutation TCIRGl（c. 1305 ＋ 2T〉C） ？TCIRGl（c. 2008OT） and CLCN7（c. 11160T） ;the case 2 carried a rare muta-tion CLCN7（c. 8570 A） . The bioinformatics analysis indicated that the rare mutations carried by these cases all had different degrees of influence on the structure and function of gene products. Conclusion The whole exome sequencing can once screen the known pathogenic mutations of osteopetrosis , is an effective tool for pathogenic mutation screening of osteopetrosis , the clinical disease in 2 cases of osteopetrosis may be closely related with the patients carrying TCIRG1 and CLCN7 mutation.