通过高通量全基因组测序技术精确判断平衡易位断点

Determination of the precise breakpoints of balanced translocation carrier by high-throughput whole genome sequencing

目的精确判定猫叫综合征患儿及其父亲(携带染色体平衡易位)的染色体异常及断点。方法通过低深度高通量全基因组测序(WGLCS,Whole-Genome Low-Coverage Sequencing)技术对患儿及其父进行测序,分析染色体中存在的染色体变异及其断点候选区域(精确的±1kb),结合PCR扩增和一代测序技术,精确确定染色体断点。结果通过WGLCS技术,将染色体断点定位到5号染色体的23,314,205到23,314,785之间和12号染色体的14,779,615到14,780,233之间。结合PCR和一代测序技术,最终将染色体平衡易位的断点确定为5号染色体的23,314,435到23,314,436之间(5p14.2)和12号染色体的14,780,019到14,780,020之间(12p13.1)。结论 WGLCS适用于染色体复杂结构变异的检测,可以精确的将染色体断点精确的±1kb区域,满足常规分子生物学的实验要求,为改进、完善人工辅助生殖技术提供可能。

Objective：Determination of the precise breakpoints of balanced translocation carrier whose child suffering from cri-du-chat syndrome;Methods：By using the whole-genome low-coverage sequencing（WGLCS）,We investigated the candidate chromosome structure variation and candidate breakpoints regions,by the combination of PCR and Sanger sequencing,we determined the precise breakpoints;Results：By using the WGLCS for each subject in this family,the 2 breakpoint regions were directly limited between 23,314,205 and 23,314,785 of chromosome 5 and between 14,779,615 and 14,780,233 of chromosome 12,with aid of PCR and Sanger sequencing,the breakpoints were finalized limited at 23,314,435 of chromosome 5 and 14,780,019 of chromosome 12;Conclusion：WGLCS can be used to effectively investigate the chromosome structure variations with high precision which can limit the breakpoints to ±1 kb region,which satisfy the requirements of experiments in molecular biology,which open the door to improve the artificial assisted reproductive technology（ART）.