Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Mol Med Rep. 2020 Aug;22(2):1269-1276. doi: 10.3892/mmr.2020.11208. Epub 2020 Jun 3.
Chromosomal abnormalities (CAs) can cause spontaneous miscarriage and increase the incidence of subsequent pregnancy loss and other complications. Presently, CAs are detected mainly by array comparative genomic hybridization (CGH) and single nucleotide polymorphism microarrays. The present study developed a low‑coverage next‑generation sequencing method to detect CAs in spontaneous miscarriage and assess its clinical performance. In total, 1,401 patients who had experienced an abortion were enrolled in the present study and divided into two groups. In group I, 437 samples that had been previously validated by array CGH were used to establish a method to detect CAs using a semiconductor sequencing platform. In group II, 964 samples, which were not verified, were assessed using established methods with respect to clinical significance. Copy number variant (CNV)‑positive and euploidy samples were verified by array CGH and short tandem repeat profiling, respectively, based on quantitative fluorescent PCR. The low‑coverage sequencing method detected CNVs >1 Mb in length and a total of 3.5 million unique reads. Similar results to array CGH were obtained in group I, except for six CNVs <1 Mb long. In group II, there were 341 aneuploidies, 195 CNVs, 25 mosaicisms and 403 euploidies. Overall, among the 1,401 abortion samples, there were 536 aneuploidies, 263 CNVs, 34 mosaicisms, and 568 euploidies. Trisomies were present in all autosomal chromosomes. The most common aneuploidies were T16, monosomy X, T22, T15, T21 and T13. Furthermore, one tetrasomy 21, one CNV associated with Wolf‑Hirschhorn syndrome, one associated with DiGeorge syndrome and one associated with both Prader‑Willi and Angelman syndromes were identified. These four cases were confirmed by short tandem repeat profiling and array CGH. Quantitative fluorescent PCR revealed nine polyploidy samples. The present method demonstrated equivalent efficacy to that of array CGH in detecting CNVs >1 Mb, with advantages of requiring less input DNA and lower cost.
染色体异常（CAs）可导致自然流产，并增加随后妊娠丢失和其他并发症的发生率。目前，CAs 主要通过比较基因组杂交（CGH）阵列和单核苷酸多态性微阵列检测。本研究开发了一种低覆盖度的下一代测序方法来检测自然流产中的 CAs，并评估其临床性能。本研究共纳入 1401 例流产患者，并分为两组。在 I 组中，使用 437 例经 CGH 阵列验证的样本建立了一种使用半导体测序平台检测 CAs 的方法。在 II 组中，使用已建立的方法评估了 964 例未经验证的样本，以评估其临床意义。基于定量荧光 PCR，通过 CGH 和短串联重复序列分析分别验证了拷贝数变异（CNV）阳性和整倍体样本。低覆盖度测序方法检测到长度>1Mb 的 CNV，共检测到 350 万个独特读段。在 I 组中获得了与 CGH 相似的结果，除了 6 个长度<1Mb 的 CNV 外。在 II 组中，有 341 例非整倍体、195 例 CNV、25 例嵌合体和 403 例整倍体。总体而言，在 1401 例流产样本中，有 536 例非整倍体、263 例 CNV、34 例嵌合体和 568 例整倍体。所有常染色体均存在三体。最常见的非整倍体是 T16、单体 X、T22、T15、T21 和 T13。此外，还发现了一例 21 三体、一例与 Wolf-Hirschhorn 综合征相关的 CNV、一例与 DiGeorge 综合征相关的 CNV 和一例与 Prader-Willi 和 Angelman 综合征均相关的 CNV。这四个病例通过短串联重复序列分析和 CGH 阵列得到了证实。定量荧光 PCR 显示有 9 例多倍体样本。本方法在检测>1Mb 的 CNV 方面与 CGH 阵列等效，具有所需输入 DNA 量少、成本低的优点。
