Department of Obstetrics and Gynecology, Seoul National University College of Medicine Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, 03080, South Korea.
Department of Biostatistics, Seoul Metropolitan Government Seoul National University Boramae Medical Center, Seoul, South Korea.
Sci Rep. 2022 Jan 7;12(1):150. doi: 10.1038/s41598-021-02507-5.
To examine the detection performance of a peptide nucleic acid (PNA) probe-based real-time time polymerase chain reaction (PCR) assay to detect common aneuploidies. Using amniotic fluid samples, PNA probe based real-time PCR (Patio DEP Detection Kit; SeaSun Biomaterials, Korea) assay was performed. PNA probe was designed to hybridize to similar sequences located on different segments of target chromosomes (21, 18, and 13) and a reference chromosome. Amplification of target sequences and melting curve analysis was performed. When analyzing the melting curve, the ratio of the peak height of the target and reference chromosome was calculated and determined as aneuploidy if the ratio of peak height was abnormal. All the results from the PNA probe-based real-time PCR and melting curve analyses were compared to those from conventional karyotyping. Forty-two cases with common aneuploidies (24 of trisomy 21, 12 of trisomy 18, and 6 of trisomy 13) and 131 cases with normal karyotype were analyzed. When comparing the karyotyping results, the sensitivity and specificity of the PNA probe-based real-time PCR assay were both 100%. The level of agreement was almost perfect (k = 1.00). PNA real-time PCR assay is a rapid and easy method for detecting common aneuploidies.
探讨肽核酸（PNA）探针实时聚合酶链反应（PCR）检测常见非整倍体的检测性能。
采用羊水样本，应用 PNA 探针实时 PCR（Patio DEP 检测试剂盒；SeaSun Biomaterials，韩国）进行检测。PNA 探针设计为与靶染色体（21、18 和 13）和参照染色体上不同片段的相似序列杂交。对靶序列进行扩增和熔解曲线分析。在分析熔解曲线时，计算目标和参照染色体峰高的比值，如果峰高比值异常，则确定为非整倍体。将 PNA 探针实时 PCR 和熔解曲线分析的所有结果与传统核型分析的结果进行比较。对 42 例常见非整倍体（21 三体 24 例，18 三体 12 例，13 三体 6 例）和 131 例正常核型进行分析。与核型分析结果比较，PNA 探针实时 PCR 检测的灵敏度和特异性均为 100%。一致性水平几乎为完美（k=1.00）。
PNA 实时 PCR 检测是一种快速、简便的检测常见非整倍体的方法。
