Shenzhen Key Laboratory for Reproductive Immunology of Preimplantation, Shenzhen Zhongshan Institute for Reproduction and Genetics, Fertility Center, Shenzhen Zhongshan Urology Hospital, Shenzhen, Guangdong, 518045, China.
Shenzhen Key Laboratory for Reproductive Immunology of Preimplantation, Shenzhen Zhongshan Institute for Reproduction and Genetics, Fertility Center, Shenzhen Zhongshan Urology Hospital, Shenzhen, Guangdong, 518045, China.
Mol Cell Probes. 2019 Aug;46:101422. doi: 10.1016/j.mcp.2019.101422. Epub 2019 Jul 15.
The vast majority of first-trimester pregnancy losses are the consequence of numerical aberrations in fetal chromosomes, which may involve nearly all chromosomes. Although commercial probes for all chromosomes are available for multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridization (FISH) analyses, their use has rarely been reported for screening all 24 chromosomes for early fetal demise, especially by FISH. Here, we validated the ability of MLPA and FISH techniques as two low-cost aneuploidy screening methods for 24 chromosomes in 165 chorionic villus samples (CVSs). The results obtained by two methods were compared by the Chi-square test and the Kappa agreement test. Both methods gave conclusive results for all CVSs tested and showed highly consistent results (kappa = 0.890, p < 0.001). There was no statistically significant difference between the aneuploidy rate of the CVSs tested by the two methods (p = 0.180). Most of the samples showed fully concordant molecular karyotyping results (81.21%) between the two analytical methods, 10.91% had incompletely concordant results, and 7.88% had discordant results. The inconsistencies included segmental abnormalities, mosaicism, and polyploidy. Both assays used to screen 24 chromosomes were powerful techniques for detecting aneuploidy in CVSs. In terms of cost-effectiveness and diagnostic accuracy, the combination of subtelomeric (P036, P070) and centromeric (P181) MLPA assays is the better analytic strategy and follow-up analysis by FISH is recommended for MLPA-negative samples.
绝大多数的早期妊娠丢失是胎儿染色体数量异常的结果，这些异常可能涉及几乎所有的染色体。虽然商业探针可用于多重连接依赖性探针扩增（MLPA）和荧光原位杂交（FISH）分析，但它们很少用于早期胎儿死亡的所有 24 条染色体的筛查，特别是 FISH 分析。在这里，我们验证了 MLPA 和 FISH 技术作为两种低成本的 24 条染色体非整倍体筛查方法在 165 个绒毛膜绒毛样本（CVS）中的应用。通过卡方检验和 Kappa 一致性检验比较两种方法的结果。两种方法对所有检测的 CVS 都给出了明确的结果，并显示出高度一致的结果（Kappa=0.890，p<0.001）。两种方法检测的 CVS 非整倍体率之间没有统计学上的显著差异（p=0.180）。大多数样本在两种分析方法之间显示出完全一致的分子核型结果（81.21%），10.91%的样本结果不完全一致，7.88%的样本结果不一致。不一致的情况包括片段异常、嵌合体和多倍体。用于筛查 24 条染色体的两种检测方法都是检测 CVS 非整倍体的有效技术。就成本效益和诊断准确性而言，端粒（P036、P070）和着丝粒（P181）MLPA 检测的组合是更好的分析策略，对于 MLPA 阴性样本，建议进行 FISH 后续分析。
