背景:拷贝数变异测序(CNV-seq)在产前诊断中至关重要,但它在检测多倍体方面的局限性,母体细胞污染(MCC),单亲二分法(UPD)限制了其在受孕产物(POC)分析中的应用。本研究旨在探讨CNV-seq时代POCs的最佳基因检测策略。
方法:在所有4,211例自发性流产病例中进行了CNV-seq和定量荧光聚合酶链反应(QF-PCR)。比较了不同的测试策略,并提出了最优的测试策略。
结果:在4,211例中,2561(60.82%)表现出临床上显著的染色体异常。单独的CNV-seq,没有QF-PCR,可能误诊311例(7.39%),包括278多倍体,13UPD,20MCC在通过QF-PCR鉴定的20例MCC病例中,CNV-seq成功查明了13例流产的原因。此外,在QF-PCR提示多倍体的情况下,CNV-seq提高了54例(1.28%)低/超三倍体病例的诊断准确性。在比较了四种不同的策略之后,顺序方法(如有必要,用CNV-seq启动,然后进行QF-PCR)是有利的,减少约70%的成本与QF-PCR,同时保持结果的准确性。
结论:我们提出了一种初始的CNV-seq,然后在需要时进行QF-PCR,这是一种有效且具有成本效益的POC遗传分析策略。
BACKGROUND: Copy number variation sequencing (CNV-seq) is crucial in prenatal diagnosis, but its limitations in detecting polyploidy, maternal cell contamination (MCC), and uniparental disomy (UPD) restrict its application in the analysis of products of conception (POCs). This study aimed to investigate an optimal genetic testing strategy for POCs in the era of CNV-seq.
METHODS: CNV-seq and quantitative fluorescent polymerase chain reaction (QF-PCR) were performed in all 4,211 spontaneous miscarriage cases. Different testing strategies were compared and the optimal testing strategies were proposed.
RESULTS: Of the 4,211 cases, 2561 (60.82%) exhibited clinically significant chromosomal abnormalities. CNV-seq alone, without QF-PCR, might misdiagnose 311 (7.39%) cases, including 278 polyploidy, 13 UPD, and 20 MCC. In 20 MCC cases identified by QF-PCR, CNV-seq successfully pinpointed the cause of miscarriage in 13 cases. Furthermore, in cases where QF-PCR suggested polyploidy, CNV-seq improved the diagnostic accuracy in 54 (1.28%) hypo/hypertriploidy cases. After comparing four different strategies, the sequential approach (initiating with CNV-seq followed by QF-PCR if necessary) emerged as advantageous, reducing approximately 70% of the cost associated with QF-PCR while maintaining result accuracy.
CONCLUSIONS: We propose an initial CNV-seq followed by QF-PCR if needed-an efficient and cost-effective strategy for the genetic analysis of POCs.