CNV-seq结合QF-PCR在孕早期流产物检测中的临床应用价值

The clinical utility of copy number variation sequencing combined with quantitative fluorescence polymerase chain reaction in early pregnancy loss

目的:探讨拷贝数变异测序(CNV-seq)结合定量荧光聚合酶链反应(QF-PCR)在孕早期流产物遗传学检测中的临床应用价值。方法:收集2019年1月至2022年2月在广东省妇幼保健院就诊的孕早期(13周以内)孕妇流产物组织样本,通过CNV-seq结合QF-PCR检测流产物组织的染色体拷贝数变异情况,分析其中染色体拷贝数异常变异的分布及发生率并评估其临床价值。结果:4145例孕早期流产物组织样本中,65.01%(2695/4145)样本提示结果异常,包括51.97%(2154/4145)非整倍体、7.04%(292/4145)三倍体、3.98%(165/4145)部分非整倍体、0.70%(29/4145)同源单亲二倍体、0.05%(2/4145)四倍体以及1.28%(53/4145)微缺失/重复。在所有异常结果中,10M以上的大片段拷贝数异常占96.96%(2613/2695)。在1.97%(53/2695)的孤立性亚显微拷贝数异常中发现了3种复发性致病性亚显微拷贝数异常,分别是22q11.21微缺失/微重复、2qter微缺失(位于2q37.3末端区域内)。结论:CNV-seq结合QF-PCR检测分析孕早期流产物组织中染色体拷贝数变异,可为流产后精准的临床管理和再生育指导提供依据;发现3种潜在的可能与流产相关的致病性亚显微拷贝数异常。

Objective:To explore the clinical utility of copy number variation sequencing(CNV-seq)and quantitative fluorescence polymerase chain reaction(QF-PCR)in identifying chromosomal anomalies in early pregnancy loss.Methods:Miscarriage specimens were collected between January 2019 and February 2022 from women who suffered early pregnancy loss in Guangdong Women and Children Hospital.CNV-seq and QF-PCR were performed to determine the chromosomal anomalies in products of conception.The distribution and incidence of chromosomal abnormalities were analyzed and the clinical significance was further evaluated.Results:A total of 4145 miscarriage specimens were collected.Abnormal results were identified in 65.01%(2695/4145)cases,including 51.97%(2154/4145)of aneuploidy,7.04%(292/4145)of triploidy,3.98%(165/4145)of partial aneuploidy(large copy number variation),0.70%(29/4145)uniparental disomy,0.05%(2/4145)of tetraploidy,and 1.28%(53/4145)of microdeletion/microduplication.The majority(96.96%,2613/2695)of abnormal results were≥10Mb and should theoretically be detectable by traditional cytogenetic analysis.Three recurrent pathogenic CNVs(microdeletions in 22q11.21,2q37.3 and microduplication in 22q11.21)were identified in 1.97%(53/2695)isolated submicroscopic copy number variation(<10Mb).Conclusions:CNV-seq combined with QF-PCR analysis of chromosome copy number variation in products of conception at the first trimester can provide evidence for further accurate clinical management and reproductive guidance.In addition,we identified three pathogenic submicroscopic copy number variations that might be associated with abortion.