使用基于联合探针锚定聚合技术的高通量测序平台的低覆盖度全基因组测序检测自然流产组织中染色体异常的分析

Chromosomal abnormalities in spontaneous miscarriage specimens detected by combinatorial probe anchor synthesis-based high-throughput low coverage whole genome sequencing

目的探讨使用基于联合探针锚定聚合技术(cPAS)的高通量测序平台的低覆盖度全基因组测序能否用以检测自然流产组织的染色体异常情况。方法2015年9月至2017年5月在内蒙古自治区妇幼保健院共收集自然流产组织样本601例,应用荧光原位杂交技术以及基于cPAS的高通量测序平台的低覆盖度全基因组测序两种方法对组织样本的染色体异常状况分别进行检测,并对比分析。结果601例组织样本中,有1例样本的基因组DNA质量不符合要求而被剔除,另有5例行FISH失败,其余595例自然流产组织样本均完成了检测。在595例自然流产组织样本中,低覆盖度全基因组测序共检出144例(24.2%,144/595)染色体异常,其中,137例(23.0%,137/595)为染色体非整倍体,2例(0.3%,2/595)为染色体嵌合体,5例(0.8%,5/595)为拷贝数变异。荧光原位杂交技术共检出131例(22.0%,131/595)染色体异常,其中,125例(21.0%,125/595)为染色体非整倍体,2例(0.3%,2/595)为染色体嵌合体,未检测出拷贝数变异。结论使用基于cPAS的高通量测序平台的低覆盖度全基因组测序是1种可靠的能够用以检测自然流产组织染色体异常的方法,其能检测包括染色体非整倍体、染色体嵌合体以及≥5 Mb的染色体拷贝数变异等在内的多种染色体异常情况。

Objective To evaluate the application of combinatorial probe anchor synthesis(cPAS)-based high-throughput low coverage whole genome sequencing in chromosomal aberration detection in spontaneous miscarriage.Methods From September 2015 to May 2017,spontaneous miscarriage samples were collected from Inner Mongolia Maternal and Child Health Care Hospital.Those samples were further analyzed with two independent methods,fluorescence in situ hybridization(FISH)and low coverage whole genome sequencing on the BGISEQ-500 high-throughput platform.The performance of low coverage whole genome sequencing was assessed by comparing to FISH results.Results In 595 spontaneous miscarried specimens,low coverage whole genome sequencing revealed 144 cases(24.2%,144/595)chromosomal abnormalities,of which a subset of 137 cases(23.0%,137/595)were detected as aneuploidies,2 cases(0.3%,2/595)as mosaicisms and 5 cases(0.8%,5/595)as copy number variation(≥5 Mb).Conclusion cPAS-based high-throughput low coverage whole genome sequencing is a reliable method in detecting chromosomal aberrations inspontaneous abortion tissues,including chromosome aneuploidies,mosaicisms and copy number variation(≥5 Mb).